Figure 1.

TRIM37 depletion induces autophagy. (a) HepG2, HEK 293T control, and TRIM37 KD cells grown in normal growth media were collected for detection of the proteins via western blotting using the indicated antibodies. Actin beta (ACTB) was used as a loading control. (b) HepG2 control and TRIM37 KD cells were immuno-stained with anti-LC3B antibody to detect the localization of endogenous LC3B proteins using regular immuno-fluorescence microscopy. (c) TRIM37 was depleted (TRIM37 KD) in HepG2 cells stably expressing mCherry-EGFP-LC3B. Cells grown in normal growth media were incubated with LysoTracker Blue (0.2 μM) for 30 min before fixation with 4% paraformaldehyde and subsequent confocal image acquisition. (d) HEK 293T control and TRIM37 KD cells were transfected with two siRNAs for ATG7 and ATG16L1. Cells were collected after 72 h for protein detection with the indicated antibodies. (e) HepG2 control and TRIM37 KD cells were treated with CQ (10 μM) for 6 h prior to western blot detection of proteins shown. s.e. short exposure; l.e. long exposure. (f) HepG2 control and TRIM37 KD cells stably expressing mCherry-EGFP-LC3B were treated with (+) or without (-) CQ (10 μM) for 6 h before confocal image acquisition. Scale bars: 10 μm. The intensities of LC3B-II were quantified and presented as the ratio of LC3B-II:ACTB below the LC3B blots as ± SEM (n = 3) from 3 independent experiments. The LC3B-II:ACTB value in control cells was set as 1.