, and were glassglass homogenized in lysis buffer (RIPA buffer, composition: 50 mM TRIS hydrochloride pH7.5, 1 mM EGTA, 1 mM EDTA, 1% Triton X-100, 0.1% SDS, 50 mM NaF, 150 mM NaCl, 0.5% Sodium deoxycholate, 0.1% mercaptoethanol, 1mM sodium orthovanadate) [91]. After centrifugation (15,000 g for 15 min at 4°C), protein concentration was determined by BCA-Assay (Pierce, 23225), A5441), rabbit anti-ULK1 (Cell Signaling Technology, 8054), rabbit anti-p-ULK1 Ser757, vol.6888, p.1, 14203.
, Ser317 (Cell Signaling Technology, 6887; discontinued), rabbit anti-PRKAA/AMPK? (Cell Signaling Technology, 2532), rabbit anti-p-PRKAA/AMPK? Thr172 (Cell Signaling Technology, 4188), guinea pig anti-SQSTM1 (Progen, GP62-C), rabbit anti-LC3B (Cell Signaling Technology, 2775), rabbit anti-RPS6 (Cell Signaling Technology, 2217), rabbit anti
Global prevalence of chronic kidney disease -a systematic review and meta-analysis, PLoS One, vol.11, issue.7, p.158765, 2016. ,
Focusing on the glomerular slit diaphragm: podocin enters the picture, Am J Pathol, vol.160, issue.1, pp.3-5, 2002. ,
Unraveling the role of podocyte turnover in glomerular aging and injury, J Am Soc Nephrol, vol.25, issue.4, pp.707-716, 2014. ,
Deficient autophagy results in mitochondrial dysfunction and FSGS, J Am Soc Nephrol, vol.26, issue.5, pp.1040-1052, 2015. ,
Autophagy influences glomerular disease susceptibility and maintains podocyte homeostasis in aging mice, J Clin Invest, vol.120, issue.4, pp.1084-1096, 2010. ,
Autophagy in human health and disease, N Engl J Med, vol.368, issue.7, pp.651-662, 2013. ,
Mechanisms of autophagy, Annu Rev Biophys, vol.44, pp.101-122, 2015. ,
Emerging regulation and functions of autophagy, Nat Cell Biol, vol.15, issue.7, pp.713-720, 2013. ,
Autophagosome formation from membrane compartments enriched in phosphatidylinositol 3-phosphate and dynamically connected to the endoplasmic reticulum, J Cell Biol, vol.182, issue.4, pp.685-701, 2008. ,
The autophagosome: origins unknown, biogenesis complex, Nat Rev Mol Cell Biol, vol.14, issue.12, pp.759-774, 2013. ,
The ER-Golgi intermediate compartment feeds the phagophore membrane, Autophagy, vol.10, issue.1, pp.170-172, 2014. ,
LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing, Embo J, vol.19, issue.21, pp.5720-5728, 2000. ,
Guidelines for the use and interpretation of assays for monitoring autophagy, Autophagy, vol.12, issue.1, pp.1-222, 2016. ,
URL : https://hal.archives-ouvertes.fr/hal-01439629
Mechanisms of autophagy initiation, Annu Rev Biochem, vol.86, pp.225-244, 2017. ,
A protein conjugation system essential for autophagy, Nature, vol.395, issue.6700, pp.395-398, 1998. ,
Mouse Apg10 as an Apg12-conjugating enzyme: analysis by the conjugation-mediated yeast two-hybrid method, FEBS Lett, vol.532, issue.3, pp.450-454, 2002. ,
WIPI-1alpha (WIPI49), a member of the novel 7-bladed WIPI protein family, is aberrantly expressed in human cancer and is linked to starvation-induced autophagy, Oncogene, vol.23, issue.58, pp.9314-9325, 2004. ,
LC3/GABARAP family proteins: autophagy-(un)related functions, Faseb J, vol.30, issue.12, pp.3961-3978, 2016. ,
Analysis of the native conformation of the LIR/AIM motif in the Atg8/LC3/GABARAP-binding proteins, Autophagy, vol.11, issue.12, pp.2153-2159, 2015. ,
Activation of ULK kinase and autophagy by GABARAP trafficking from the centrosome is regulated by WAC and GM130, Mol Cell, vol.60, issue.6, pp.899-913, 2015. ,
The ULK1 complex: sensing nutrient signals for autophagy activation, Autophagy, vol.9, issue.2, pp.124-137, 2013. ,
The role of the Atg1/ULK1 complex in autophagy regulation, Curr Opin Cell Biol, vol.22, issue.2, pp.132-139, 2010. ,
ULK-Atg13-FIP200 complexes mediate mTOR signaling to the autophagy machinery, Mol Biol Cell, vol.20, issue.7, pp.1992-2003, 2009. ,
ATG13.FIP200 complex mediates mTOR signaling and is essential for autophagy, J Biol Chem, vol.284, issue.18, pp.12297-12305, 2009. ,
Nutrient-dependent mTORC1 association with the ULK1-Atg13-FIP200 complex required for autophagy, Mol Biol Cell, vol.20, issue.7, pp.1981-1991, 2009. ,
Impaired podocyte autophagy exacerbates proteinuria in diabetic nephropathy, Diabetes, vol.65, issue.3, pp.755-767, 2016. ,
Endothelial cell and podocyte autophagy synergistically protect from diabetes-induced glomerulosclerosis, Autophagy, vol.11, issue.7, pp.1130-1145, 2015. ,
URL : https://hal.archives-ouvertes.fr/hal-01539607
Role of mTOR in podocyte function and diabetic nephropathy in humans and mice, J Clin Invest, vol.121, issue.6, pp.2197-2209, 2011. ,
Emerging role of autophagy in kidney function, diseases and aging, Autophagy, vol.8, issue.7, pp.1009-1031, 2012. ,
mTORC1 activation in podocytes is a critical step in the development of diabetic nephropathy in mice, J Clin Invest, vol.121, issue.6, pp.2181-2196, 2011. ,
Targeting mTOR signaling can prevent the progression of FSGS, J Am Soc Nephrol, 2017. ,
mTOR signaling in growth control and disease, Cell, vol.149, issue.2, pp.274-293, 2012. ,
Dissection of the autophagosome maturation process by a novel reporter protein, tandem fluorescent-tagged LC3, Autophagy, vol.3, issue.5, pp.452-460, 2007. ,
New autophagy reporter mice reveal dynamics of proximal tubular autophagy, J Am Soc Nephrol, vol.25, issue.2, pp.305-315, 2014. ,
Autophagy in the liver, J Hepatol, vol.59, issue.2, pp.389-391, 2013. ,
The role of autophagy in liver diseases: mechanisms and potential therapeutic targets, Biomed Res Int, p.480508, 2015. ,
Methods in mammalian autophagy research, Cell, vol.140, issue.3, pp.313-326, 2010. ,
Guidelines for the use and interpretation of assays for monitoring autophagy, Autophagy, vol.8, issue.4, pp.445-544, 2012. ,
URL : https://hal.archives-ouvertes.fr/hal-01439629
Podocyte-specific expression of cre recombinase in transgenic mice, Genesis, vol.35, issue.1, pp.39-42, 2003. ,
In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker, Mol Biol Cell, vol.15, issue.3, pp.1101-1111, 2004. ,
Podocyte hypertrophy, "adaptation," and "decompensation" associated with glomerular enlargement and glomerulosclerosis in the aging rat: prevention by calorie restriction, J Am Soc Nephrol, vol.16, issue.10, pp.2953-2966, 2005. ,
PRAS40 acts as a nodal regulator of high glucose-induced TORC1 activation in glomerular mesangial cell hypertrophy, J Cell Physiol, vol.225, issue.1, pp.27-41, 2010. ,
p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death, J Cell Biol, vol.171, issue.4, pp.603-614, 2005. ,
AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1, Nat Cell Biol, vol.13, issue.2, pp.132-141, 2011. ,
Phosphorylation of ULK1 (hATG1) by AMP-activated protein kinase connects energy sensing to mitophagy, Science, vol.331, issue.6016, pp.456-461, 2011. ,
The autophagy initiating kinase ULK1 is regulated via opposing phosphorylation by AMPK and mTOR, Autophagy, vol.7, issue.6, pp.643-644, 2011. ,
Pharmacological inhibition of ULK1 kinase blocks mammalian target of rapamycin (mTOR)-dependent autophagy, J Biol Chem, vol.290, issue.18, pp.11376-11383, 2015. ,
The contribution of chronic kidney disease to the global burden of major noncommunicable diseases, Kidney Int, vol.80, issue.12, pp.1258-1270, 2011. ,
Has the incidence of end-stage renal disease in the USA and other countries stabilized?, Curr Opin Nephrol Hypertens, vol.20, issue.3, pp.241-245, 2011. ,
Prevalence and socio-economic aspects of chronic kidney disease, Nephrol Dial Transplant, vol.17, issue.11, pp.8-12, 2002. ,
Transgenic expression of human APOL1 risk variants in podocytes induces kidney disease in mice, Nat Med, vol.23, issue.4, pp.429-438, 2017. ,
Autophagosome maturation is impaired in Fabry disease, Autophagy, vol.6, issue.5, pp.589-599, 2010. ,
Disruption of the beclin 1-BCL2 autophagy regulatory complex promotes longevity in mice, Nature, vol.558, issue.7708, pp.136-140, 2018. ,
Overexpression of Atg5 in mice activates autophagy and extends lifespan, Nat Commun, vol.4, p.2300, 2013. ,
Tor, a phosphatidylinositol kinase homologue, controls autophagy in yeast, J Biol Chem, vol.273, issue.7, pp.3963-3966, 1998. ,
Role and regulation of starvation-induced autophagy in the Drosophila fat body, Dev Cell, vol.7, issue.2, pp.167-178, 2004. ,
Nutrient starvation elicits an acute autophagic response mediated by Ulk1 dephosphorylation and its subsequent dissociation from AMPK, Proc Natl Acad Sci, vol.108, issue.12, pp.4788-4793, 2011. ,
Spatial coupling of mTOR and autophagy augments secretory phenotypes, Science, vol.332, issue.6032, pp.966-970, 2011. ,
Relieving autophagy and 4EBP1 from rapamycin resistance, Mol Cell Biol, vol.31, issue.14, pp.2867-2876, 2011. ,
Role of AMPK-mTOR-Ulk1/2 in the regulation of autophagy: cross talk, shortcuts, and feedbacks, Mol Cell Biol, vol.32, issue.1, pp.2-11, 2012. ,
mTOR: a pharmacologic target for autophagy regulation, J Clin Invest, vol.125, issue.1, pp.25-32, 2015. ,
, Mol Cell Biol, vol.31, issue.15, pp.3082-3084, 2011.
mTOR and autophagy: a dynamic relationship governed by nutrients and energy, Semin Cell Dev Biol, vol.36, pp.121-129, 2014. ,
The AMPK signalling pathway coordinates cell growth, autophagy and metabolism, Nat Cell Biol, vol.13, issue.9, pp.1016-1023, 2011. ,
AMPK phosphorylation of raptor mediates a metabolic checkpoint, Mol Cell, vol.30, issue.2, pp.214-226, 2008. ,
ULK1 inhibits mTORC1 signaling, promotes multisite Raptor phosphorylation and hinders substrate binding, Autophagy, vol.7, issue.7, pp.737-747, 2011. ,
Ulk1-mediated phosphorylation of AMPK constitutes a negative regulatory feedback loop, Autophagy, vol.7, issue.7, pp.696-706, 2011. ,
Inhibition of MTOR disrupts autophagic flux in podocytes, J Am Soc Nephrol, vol.23, issue.3, pp.412-420, 2012. ,
MTOR regulates autophagic flux in the glomerulus, Autophagy, vol.8, issue.4, pp.696-698, 2012. ,
Lysosomal mTORC2/PHLPP1/Akt regulate chaperone-mediated autophagy, Mol Cell, vol.59, issue.2, pp.270-284, 2015. ,
Plasma membrane stress induces relocalization of Slm proteins and activation of TORC2 to promote sphingolipid synthesis, Nat Cell Biol, vol.14, issue.5, pp.542-547, 2012. ,
Sustained activation of mTORC1 in macrophages increases AMPKalpha-dependent autophagy to maintain cellular homeostasis, BMC Biochem, vol.17, issue.1, p.14, 2016. ,
The rapamycin-sensitive complex of mammalian target of rapamycin is essential to maintain male fertility, Am J Pathol, vol.186, issue.2, pp.324-336, 2016. ,
Post-rapamycin proteinuria: incidence, evolution, and therapeutic handling at a single center, Transplant Proc, vol.38, issue.10, pp.3476-3478, 2006. ,
Post-transplantation proteinuria and sirolimus, N Engl J Med, vol.353, pp.2088-2089, 2005. ,
Clinically significant proteinuria following the administration of sirolimus to renal transplant recipients, Drug Metab Lett, vol.1, issue.4, pp.267-271, 2007. ,
Adverse events associated with mTOR inhibitors, Expert Opin Drug Saf, vol.12, issue.2, pp.177-186, 2013. ,
High sirolimus levels may induce focal segmental glomerulosclerosis de novo, Clin J Am Soc Nephrol, vol.2, issue.2, pp.326-333, 2007. ,
Adipose-specific knockout of raptor results in lean mice with enhanced mitochondrial respiration, Cell Metab, vol.8, issue.5, pp.399-410, 2008. ,
URL : https://hal.archives-ouvertes.fr/inserm-00351013
Critical role for hypothalamic mTOR activity in energy balance, Cell Metab, vol.9, issue.4, pp.362-374, 2009. ,
VEGF inhibition and renal thrombotic microangiopathy, N Engl J Med, vol.358, issue.11, pp.1129-1136, 2008. ,
Suppression of basal autophagy in neural cells causes neurodegenerative disease in mice, Nature, vol.441, issue.7095, pp.885-889, 2006. ,
Systemic treatment with the antidiabetic drug metformin selectively impairs p53-deficient tumor cell growth, Cancer Res, vol.67, issue.14, pp.6745-6752, 2007. ,
Clinical pharmacokinetics of sirolimus, Clin Pharmacokinet, vol.40, issue.8, pp.573-585, 2001. ,
N-wasp is required for stabilization of podocyte foot processes, J Am Soc Nephrol, vol.24, issue.5, pp.713-721, 2013. ,
A global double-fluorescent Cre reporter mouse, Genesis, vol.45, issue.9, pp.593-605, 2007. ,
Quantitative imaging of basic functions in renal (patho)physiology, Am J Physiol Renal Physiol, vol.291, issue.2, pp.495-502, 2006. ,
Tracking the fate of glomerular epithelial cells in vivo using serial multiphoton imaging in new mouse models with fluorescent lineage tags, Nat Med, vol.19, issue.12, pp.1661-1666, 2013. ,
Large-scale identification of genes implicated in kidney glomerulus development and function, Embo J, vol.25, issue.5, pp.1160-1174, 2006. ,
Molecular fingerprinting of the podocyte reveals novel gene and protein regulatory networks, Kidney Int, vol.83, issue.6, pp.1052-1064, 2013. ,
URL : https://hal.archives-ouvertes.fr/hal-01001262
A new method for large scale isolation of kidney glomeruli from mice, Am J Pathol, vol.161, issue.3, pp.799-805, 2002. ,
A conditionally immortalized human podocyte cell line demonstrating nephrin and podocin expression, J Am Soc Nephrol, vol.13, issue.3, pp.630-638, 2002. ,
HK-2: an immortalized proximal tubule epithelial cell line from normal adult human kidney, Kidney Int, vol.45, issue.1, pp.48-57, 1994. ,