Fig. 4: PEC inhibits the autophagic flux in BLCA cells.

A Western blots showing protein levels of p62 and LC3B after incubation with PEC for 24 h. B Representative TEM images of BLCA cells exposed to PEC (10 μM in 5637, 20 μM in SCaBER) for 24 h. The red arrows mark the autophagic vacuole. Scale bar, 1 μm. C Western blots showing estimation of levels of p62 and LC3B in BLCA cells treated with PEC (10 μM in 5637, 20 μM in SCaBER) for 24 h and with CQ (100 μM) in the last 6 h. D Western blot analysis of p62 and LC3B in BLCA cells incubated with PEC (10 μM in 5637, 20 μM in SCaBER) for 24 h or CQ (100 μM) in the last 6 h or combination with serum starvation. E Immunofluorescence analysis of p62 and LC3B colocalization in GFP-LC3-Hela cells incubated with PEC (20 μM) or CQ (100 μM) in the last 6 h or combination. Scale bar, 10 μm. F Immunofluorescence analysis of mCherry-EGFP-LC3B 5637 stable cell line treated with PEC (10 μM, 24 h) or CQ (100 μM, 6 h). Scale bar, 10 μm. G MTT assay data showing enhancement of the inhibitory effect of PEC (10 μM in 5637, 20 μM in SCaBER) in BLCA cells upon ATG7 knockdown by 48 h. H LC3B and γ-H2AX protein levels in BLCA cells transfected with siATG7 for 48 h, followed by treatment with or without PEC (10 μM in 5637, 20 μM in SCaBER) for 24 h. I Western blots showing autophagy-related proteins in BLCA cells after knocking down TOP2A or PEC treatment (10 μM in 5637, 20 μM in SCaBER) or combination.