Fig. 3

Talin2 functionally interacts with KANK2. A Knockdown of talin1, unlike talin2, leads to changes in KANK2 localization. Forty-eight hours after transfection with either control siRNA, talin1 or talin2-specific siRNA, MDA-MB-435S cells were methanol fixed and stained with anti-talin1 antibody followed by Alexa-Fluor IgG1 555-conjugated antibody (red), anti-talin2 antibody followed by Alexa-Fluor IgG2b 488-conjugated antibody (green), anti-KANK2 antibody followed by Alexa-Fluor 647-conjugated antibody (magenta) and IRM images were taken. Analysis was performed using TCS SP8 Leica. Scale bar = 10 µm. B Quantification of data presented in (A). Violin plot represents measurements of > 30 cells. Data were analyzed by one-way ANOVA with Dunnett’s multiple comparison. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. C WB analysis of talin1, talin2, liprin-β1, KANK2 or integrin β5 in IACs isolated from MDA-MB-435S cells transfected with either control siRNA, talin1 or talin2-specific siRNA. Forty-eight hours after transfection, IACs were isolated and WB analysis was performed. The results presented are representative of two independent experiments yielding similar results. Numbers below WB scan represent expression of target proteins after talin1 or talin2 knockdown as relative to expression of target proteins in cell transfected with control siRNA set as 1