Quantification of PrPC in normal N2a58 cells and PrPSc in prion infected N2a58/22L cells after 3 and 6 days of treatment with antibodies
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Quantification of PrPC in normal N2a58 cells and PrPSc in prion infected N2a58/22L cells after 3 and 6 days of treatment with antibodies
| Treatment of cell with Antibodiesa | |||||||
|---|---|---|---|---|---|---|---|
| Incubation(days) | Control | SAF34 (1 µg/mL) | SAF61 (1 µg/mL) | Expected values if additive effects | SAF34 + SAF61 (1 µg/mL each) | Statistical analysis | |
| PrPC | 3 days | 100b | 90 ± 1 | 67 ± 8 | 57 ± 9 | 33 ± 3 | p < 0.001e |
| 6 days | 100 | 84 ± 4 | 43 ± 6 | 27 ± 10 | 23 ± 4 | pp > 0.05 | |
| PrPSc | 3 days | 100c | 80 ± 4 | 43 ± 3 | 23 ± 7 | 9 ± 0.7 | p < 0.01 |
| 6 days | 100 | 35 ± 2 | < 1d | – | < 1d | – | |
bPrPC. bands were quantified using the image analysis software Scions Image Beta 4.02. Values are given as densitometry units (DU) where 100% corresponds to the intensity of the PrPC band in the absence of antibody and 0% represents undetectable levels of PrPC in the immunoblot. For each PrPC band, we subtracted a background value, selected in its vicinity. All the densitometry values were normalized in comparison with their respective controls. Data represent the mean from three independent experiments. Calculated errors correspond to the standard deviation.
eStatistical analyses were carried out using the Graph Pad Prism software (Graph Pad Software, San Diego, USA). Values were analysed by anova followed by the Newman·Keuls multiple comparison test (n = 3). First we investigated whether the values obtained after treatment with SAF34, SAF61 and both antibodies were statistically significant compared with untreated control cells (treated cells vs. control, p < 0.001). Second we compared the experimental values obtained when cells were co-treated with both SAF34 and SAF61 antibodies versus the values expected in the case of additive effects. The result of this second analysis is only mentioned in the last column of the Table 2.