Supporting data for 'Kindlin-driven inside-out activation governs force-dependent adhesion assembly of integrin beta6'

Super-resolution fluorescence microscopy was utilized to examine the spatiotemporal regulation of integrin beta6 on RGD-membrane devoid of traction force. RT-qPCR has revealed a high abundance of talin1, kindlin1 and kindlin2 in CHO-B2 cells. However, solely kindlin2 critically reinforced the adhesion clustering between RGD ligands and integrin beta6. When integrin beta6 cytoplasmic tail was swapped with that of integrin beta1, denser RGD clustering was observed via the pixel-intensity image analysis. To that end, we conducted biochemistry assay (immunoprecipitation and western blot), and ultimately demonstrated a preferential recruitment of kindlin2 by integrin beta1 comparing to integrin beta6. Kindlin2 introduction also certainly enhanced integrin beta6 activation and strengthened the adhesion formation on RGD-membrane. Remarkably, additional kindlin2 was found to support cell locomotion even on the soft substrate, thereby overcame the rigidity-dependent migration of integrin beta6.