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. Author manuscript; available in PMC: 2024 Mar 16.
Published in final edited form as: Mol Cell. 2023 Mar 9;83(6):927–941.e8. doi: 10.1016/j.molcel.2023.02.023

Fig. 3. TNIP1 is a negative regulator of selective autophagy.

Fig. 3

(A) IB of HeLa cells, HeLa TNIP1 KO cells and HeLa TNIP1 KO rescued with FKBP-GFP-TNIP1. Green arrow, FKBP-GFP-TNIP1 construct. Black arrow, endogenous TNIP1.

(B) HeLa cells stably expressing mito-mKeima and HA-Parkin in wild type cells, TNIP1 KO cells or TNIP1 KO cells rescued with FKBP-GFP-TNIP1 construct were treated with Oligomycin and Antimycin (O/A) for 6 h and subjected to FACS analysis. Left, representative FACS plot. Right, bar graph representing data as mean ± SEM obtained from 3 independent replicates.

(C) HeLa cells stably expressing mito-mKeima and HA-Parkin in wild type cells, TNIP1 KO cells or TNIP1 KO cells rescued with FKBP-GFP-TNIP1 construct were treated with O/A for 2 h and subjected to FACS analysis. Left, representative FACS plot. Right, bar graph representing data as mean ± SEM obtained from 3 independent replicates.

(D) HeLa cells stably expressing mito-mKeima, HA-Parkin and overexpressing FKBP-GFP-TNIP1 wild type, ΔAHD1, ΔAHD3 or ΔAHD4 constructs were treated with O/A for 2 h and subjected to FACS analysis. Left, representative FACS plot. Right, bar graph representing data as mean ± SEM obtained from 3 independent replicates.

(E) HeLa TNIP1 KO cells stably expressing mito-mKeima, HA-Parkin and rescued with FKBP-GFP-TNIP1 wild type, LIR2 mutant or ΔAHD3 constructs were treated with O/A for 3 h and subjected to FACS analysis. Left, representative FACS plot. Right, bar graph representing data as mean ± SEM obtained from 3 independent replicates.

(F) IB of HeLa cells stably expressing BFP-Parkin and GFP-TNIP1 WT treated for 15 h, 18 h, 21 h or 24 h with O/A. Right, bar graph representing data as mean ± SEM obtained from 3 independent replicates.