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, WM3912 (middle) and WM8 (right) melanoma cells transfected with a control (siCtl) or two different ASAH1 (siASAH1#1 and siASAH1#2) siRNA. The proform (55 kDa) and the ? (40 kDa) active subunit are shown. The ? (14 kDa) active subunit is not recognized by the commercial antibody
, ASAH1 activity in the corresponding melanoma cells. Values represent mean + SD of two independent experiments
, TEM images of WM8 melanoma cells transfected with control or ASAH1 siRNA. Scale bars, vol.5
, Colony formation assay of melanoma cells transfected with control, p.1
Colonies were stained with crystal violet after 14 days (representative micrographs are shown) and destained. The relative absorbance, which reflects the number of colony was measured on a spectrophotometer. Values represent mean + SD of three independent experiments ,
, , p.1
, WM3912 (middle) and WM8 (right) melanoma cells transfected with a control (siCtl), two ASAH1 (siASAH1#1 and siASAH1#2) or MITF (siMITF) siRNA. DAPI positive cells represent dying cells, FACS analysis of DAPI staining in 501mel (left)
, Figure S4: ASAH1 controls melanoma cell proliferation (A) Immunoblot to ASAH1 of WM3918 melanoma cell lines with forced expression of
, or ITG?V?5 neutralizing antibody (ITG?V?5) the last 5 hours. Cells positive for DAPI represent dying cells. Results are expressed in %
, Boyden chamber assays with WM8 and WM3912 melanoma cells transfected with control, ASAH1 or MITF siRNA in presence of control IgG (Ctl) or ITG?V?5 neutralizing antibody. Values represent mean + SD of three independent experiments