Fig. 3.

A dual role for autophagy in HOG-LDL-induced pericyte death. (a–c) HRCPs were treated as follows: exposure to N-LDL (300 mg/l) or HOG-LDL (25–300 mg/l) for 24 h (a); pre-treatment with 3-MA (5 mmol/l, light grey bars), Z-VAD-fmk (100 μmol/l, dark grey bars) and both of them (black bars) for 1 h (b); transfection with siRNA against Beclin-1 (si-Beclin-1, black bars) or si-Ctrl (white bars) for 36 h, then exposure to HOG-LDL (50 mg/l, 200 mg/l) for 12 h (c). Cell viability was expressed as percentage vs control. (d-f) HRCPs: apoptosis and autophagy. Apoptosis is triggered by HOG-LDL at 100 or 200 mg/l: cleaved PARP (white bars), activated caspase-3 (black bars) were detected by western blot (d). HRCPs were pre-treated with 3-MA (5 mmol/l) or CQ (10 μmol/l) for 1 h: cleaved PARP (white bars) and activated caspase-3 (black bars) were detected by western blot (e). HRCPs were pre-treated with vehicle (white bars) or 3-MA (5 mmol/l, grey bars) for 1 h, or were transfected with si-Beclin-1 (black bars), then exposed to HOG-LDL (50 mg/l, 200 mg/l) for 12 h: TUNEL staining was expressed as a ratio of control (f). All data are means ± SD, n = 3 or 5; *p < 0.05, **p < 0.01. Ctrl, control