Fig. 5. ATG4B plays an important role in ChC^ILpre-exos-mediated inhibition of autophagy.

a RT-PCR was used to detect the mRNA change of autophagy-related gene in PMA-induced THP-1 cells with the indicated treatments. b PMA-induced THP-1 cells were given with the indicated treatments for 12 h and then the ATG4B protein level was detected by western blot. c PMA-induced THP-1 cells were preincubated with 10 µM CHX for the indicated time, and then incubated swChC^NCpre-exos or swChC^ILpre-exos in the presence of LPS for an additional 12 h. Subsequently, the ATG4B protein level was evaluated by western blot. d PMA-induced THP-1 cells were transfected with Lenti-ATG4B shRNA or its control shRNA. Then the protein levels of ATG4B and LC3I/II were detected by western blot. e, f PMA-induced THP-1 cells were treated as d and then the immunofluorescence assay was taken to measure the LC3-puncta number per cell (e) and the ratio of ASC-specking cells (f). ANOVA with Bonferroni’s multiple comparison test was used, ***p < 0.001; *p < 0.05; ns, no significance