TRIM27 cooperates with STK38L to inhibit ULK1‐mediated autophagy and promote tumorigenesis

A

Verification of ULK1 knockout in HeLa cells by Western blot.

B

p62 or LC3 levels in HeLa ULK1 KO cells co‐expressing vector control, HA‐ULK1 WT, or HA‐ULK1 K568/K571R.

C

Quantification of LC3‐II/LC3‐I ratio, and LC3‐II/actin ratio with Bafilomycin A1 treatment related to Fig 3C. Data are mean ± SEM for three independent experiments. P‐values were determined by an unpaired t‐test with Holm‐Sidak’s method for multiple t‐tests, ***P < 0.001.

D

Representative images of GFP fluorescence localization in HeLa ULK1 KO cells co‐expressing GFP‐LC3 together with vector control, HA‐ULK1 wildtype, or HA‐ULK1 K568/K571R, respectively.

E

Quantitation of data presented in (D) shows mean number of GFP‐LC3 puncta per cell ± SD (n = 30 cells per condition). P‐values were determined by a one‐way ANOVA with Tukey’s multiple comparisons test, ***P < 0.001.

F, G

Western blot analysis of ULK1 accumulation and LC3 conversion in the liver of 3‐month‐old Trim27^+/+ and Trim27^−/− mice. Each lane represents a different mouse (F). Quantification of ULK1, LC3‐II/LC3‐I, and LC3‐II/actin ratios in F (G).

H, I

Western blot analysis of ULK1 accumulation and LC3 conversion in the spleen of 3‐month‐old Trim27^+/+ and Trim27^−/− mice. Each lane represents a different mouse (H). Quantification of ULK1, LC3‐II/LC3‐I, and LC3‐II/actin ratios in H (I).

J, K

Western blot analysis of ULK1 accumulation and LC3 conversion in the brain of 3‐month‐old Trim27^+/+ and Trim27^−/− mice. Each lane represents a different mouse (J). Quantification of ULK1, LC3‐II/LC3‐I, and LC3‐II/actin ratios in J (K).

L, M

Western blot analysis of ULK1 accumulation and LC3 conversion in the thymus of 3‐month‐old Trim27^+/+ and Trim27^−/− mice. Each lane represents a different mouse (L). Quantification of ULK1, LC3‐II/LC3‐I, and LC3‐II/actin ratios in L (M).

Figure EV2. Knockout of TRIM27 leads to an increase of autophagy in multiple tissues.