Figure 6: Downregulation of CED components promotes apoptosis of the cells induced by accumulation of stably expressed misfolded CFTR-ΔF508.
From: Misfolded polypeptides are selectively recognized and transported toward aggresomes by a CED complex
(a) TUNEL assay of HeLa cells stably expressing misfolded protein CFTR-ΔF508. HeLa cells were transfected with the indicated siRNA. Two days later, the cells were untreated or treated with MG132 for 12 h, and then analysed by a TUNEL assay (Supplementary Fig. 11). The ratio of stained cells was calculated and is represented as a percentage. (b–d) Complementation assay using siRNA-resistant (R) CTIF variants. HeLa cells stably expressing misfolded protein CFTR-ΔF508 were transfected with either CTIF siRNA or nonspecific control siRNA. One day later, cells were re-transfected with a plasmid expressing FLAG-CTIFR-WT or FLAG-CTIFR(54–598). One day later, the cells were treated with MG132 for 12 h, and then analysed by western blotting (b) and a TUNEL assay (d). Scale bar, 50 μm. The ratio of stained cells was calculated and is represented as a percentage (c). n=3, *P<0.05; **P<0.01.
