Fig. 6: LIM domain deletion analysis by photoactivation and FA-isolation.

a Schematic representation of the paxillin LIM domain deletion mutants. All proteins were N-terminally tagged with PA-GFP or mCherry. b Alignment of the four paxillin LIM domains. Gray: zinc-binding residues. c Western blot of protein extracts from NIH-3T3 fibroblasts transiently co-expressing β3^WT_GFP and mCherry-tagged paxillin proteins, wt or deletion mutants. Loading control: Ponceau staining. d Experimental dissociation kinetics of PA-GFP_paxillin wt and deletion mutants from β3^WT_mCherry-positive FAs. e Box plot of the corresponding half-lives Statistical analysis is provided in Supplementary Dataset 4. f Quantification of mCherry_paxillin wt and mutants retained in FAs isolated from NIH-3T3 cells co-expressing β3^WT_GFP. Statistical analysis is provided in Supplementary Dataset 1. g Representative TIRF images of FAs in NIH-3T3 cells co-expressing β3^WT_GFP and mCherry_paxillin proteins. Left panel: FAs in intact cells; right panel: isolated FAs. ns (not significant) p > 0.05; *p ≤ 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. Sample size, complete statistical analysis, and p values are provided in tables in supplementary datasets.