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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. 1. Ectopic localization of TNIP1 to the mitochondria induces mitophagy.

Fig. 1

(A) Schematic representation of TNIP1 and its 2 identified putative LIR motifs.

(B) HeLa cells stably expressing HA-TNIP1 wild type, LIR1 mutant, LIR2 mutant or LIR1+LIR2 double mutant co-immunoprecipitated (co-IP) with purified recombinant GST tagged mATG8 proteins and subjected to immunoblot analysis (IB).

(C) Schematization of CID experiment.

(D) HeLa cells stably expressing mito-mKeima, FRB-FIS1 and FKBP-GFP-TNIP1 WT or UBAN mutant (D472N) were treated with Rapalog for 24 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 cells stably expressing mito-mKeima, FRB-FIS1 and FKBP-GFP-TNIP1 WT, LIR1 mutant, LIR2 mutant or LIR1&LIR2 double mutant were treated with Rapalog for 24 h and subjected to FACS acquisition. Left, representative FACS plot. Right, bar graph representing data as mean ± SEM obtained from 3 independent replicates.

(F) Top, schematic representation of the TNIP family TNIP1, TNIP2 and TNIP3. Bottom, HeLa cells stably expressing mito-Keima, FRB-FIS1 and FKBP-GFP-TNIP1, FKBP-GFP-TNIP2 or FKBP-GFP-TNIP3 were treated with Rapalog for 24 h and subjected to FACS acquisition. Left, representative FACS plot. Right, bar graph representing data as mean ± SEM obtained from 3 independent replicates.