Figure 4
From: NDRG1 inhibition sensitizes osteosarcoma cells to combretastatin A-4 through targeting autophagy
NDRG1 knockdown inhibits autophagosome-lysosome fusion in OS cells. (a) MG63.2 cells were transfected with control nontarget or NDRG1 siRNA, and the whole-cell lysates were subjected to immunoblotting. The lower panel showed quantitation of NDRG1 double bands. (b) MG63.2 cells transfected with control nontarget or NDRG1 siRNA either cultured under free-serum starved or CQ treatment, and the whole-cell lysates were subjected to immunoblotting of LC3B-II and GAPDH. (c) The LC3-II bands in (b) were quantified and normalized according GAPDH. (d and e) MG63.2 cells stably expressing the mCherry-GFP-LC3 reporter were transfected with control nontarget or NDRG1 siRNA. The colocalizations of mCherry and GFP puncta were examined by the confocal microscopy. (f) and (g) NDRG1 knockdown induced the accumulation of AVs as shown in the electron micrographs. The arrow indicates AVs. (h) Control nontarget and NDRG1 siRNA treated MG63.2 cells stably expressing GFP-LC3, and stained with antibodies against LAMP1 for confocal microscopy. (i) Quantification of average GFP-LC3 puncta per cell. (j) Quantification of GFP-LC3/LAMP1 colocalization co-efficiency. The data were presented as mean±S.D (*P<0.05, n=3)
