1. TRIM17 and Beclin 1 colocalize with the anti-autophagy factor Mcl-1.
TRIM17 and Beclin 1 colocalize with the anti-autophagy factor Mcl-1. (A) Confocal microscopic analysis of the GFP–TRIM17 and LAMP2 localization in HeLa cells. Arrows indicate colocalizing puncta. (B) High-content imaging analysis of TFEB nuclear translocation. Left, representative image of HeLa cells subjected to TRIM17 knockdown (siTRIM17.1), treated with pp242, and stained to detect TFEB and DNA. The bottom image shows the cell boundary (whites line) and TFEB-positive nuclei (yellow line). Right, the percent of nuclei showing TFEB-positivity under the indicated conditions. siScr, scrambled siRNA. Data are means±s.e.m., n=3 experiments. †P>0.05 (ANOVA). (C) Co-immunoprecipitation (IP) analysis of interactions between TRIMs and active (pSer317) versus inactive (pSer757) phospho-ULK1 from the lysates of transfected HEK293T cells as assessed by western blotting (WB). (D) Confocal microscopic analysis of the GFP–TRIM17 and FLAG–Beclin-1 localization in HeLa cells. Arrows indicate colocalizing puncta. (E) Confocal microscopic analysis of the GFP–TRIM17 and FLAG–Mcl-1 localization in HeLa cells. (F) Confocal microscopic analysis of the mCherry–TRIM17, Beclin-1–GFP and FLAG–Mcl-1 localization in transfected HeLa cells. Arrows indicate colocalizing puncta. Enlargements of the indicated area in A, D E and F are shown on the right of the main images.
Michael A. Mandell et al. J Cell Sci 2016;129:
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