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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. 5. TNIP1 binds FIP200 via its LIR motif and to the CLAW domain of FIP200.

Fig. 5

(A) Co-IP and IB of HEK293T cells stably expressing HA-TNIP1 full length or the ΔAHD1, ΔAHD3, ΔAHD4 and LIR2 mutant constructs using magnetic HA beads.

(B) HeLa wild type, TAX1BP1 KO and LC3/GABARAP 6KO cells stably expressing FRB-FIS1 and FKBP-GFP-TNIP1 WT or LIR2m were treated for 24 h with Rapalog and stained for endogenous FIP200 before immunofluorescence acquisition on a confocal microscope. Quantifications as mean ± SEM of Pearson correlation coefficient representing colocalization between FIP200 and GFP. See Fig S5B for representative images.

(C) Top, schematic representation of full length FIP200 and the regions encompassing the N-terminal and C-terminal domains as well as the minimal leucine zipper (LZ) and Claw (CLW) domains. Bottom, Co-IP and IB of HEK293T cells stably expressing GFP-TNIP1 full length and transiently expressing HA-FIP200 full length, N-terminal, C-terminal, LZ and CLW constructs using magnetic GFP beads. The zone in dotted lines was exposed longer to reveal the binding to the CLAW domain (lower part).