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. 2019 Sep 25;3(11):1520–1543. doi: 10.1002/hep4.1429

Figure 1.

Figure 1

SULF2 induces autophagic flux. (A) Western blot analysis shows protein‐expression changes of LC3B conversion and p62 levels according to SULF2 status in Huh7 and Hep3B cells in the absence or presence of bafilomycin‐A1. (B) Confocal microscopic images shows GFP‐LC3 puncta according to SULF2 status in Huh7 and Hep3B cells in the absence or presence of bafilomycin‐A1. (C) Tandem RFP‐GFP‐LC3B assay shows autophagosomes (RFP+/GFP+, yellow puncta) and autolysosomes (RFP+/GFP‐, red puncta) according to SULF2 status in Huh7 and Hep3B cells in the absence or presence of bafilomycin‐A1. Yellow bar indicates autophagosomes and red bar indicates autolysosomes. (D) Ultrastructural evidence of autophagy according to SULF2 status in Huh7 and Hep3B cells in the absence or presence of bafilomycin‐A1. Yellow arrows indicate autophagosomes and red arrows indicate autolysosomes (scale bars: 500 nm). Bafilomycin‐A1 (100 nM) was treated to inhibit the fusion between autophagosomes and lysosomes. Quantification was performed by counting a total of 50 cells in 10 random fields (5 cells/field) and presented as bar graphs (mean ± SEM). Abbreviations: GFP, green fluorescent protein; RFP, red fluorescent protein; SEM; standard error of the mean; SULF2, sulfatase 2.