Fig. 1: Force-FAK localization and force-pY397-FAK are linearly coupled at individual FAs. | Nature Communications

Fig. 1: Force-FAK localization and force-pY397-FAK are linearly coupled at individual FAs.

From: Force-FAK signaling coupling at individual focal adhesions coordinates mechanosensing and microtissue repair

Fig. 1

a Schematic of individual FA traction force measurement for a cell using mPADs. Fluorescence images showing (b) mPAD posts, (c) tFAK, (d) pY397-FAK, and (e) eGFP-vinculin for cell on mPAD. f–i Heatmaps for (f, h) tFAK and (g, i) pY397-FAK staining at FAs on mPADs of 5 or 14 kPa. White arrows indicate force magnitudes and directions. Scale bar 5 µm. j Force (mean ± SD) at individual FAs on 5 kPa mPADs. Two-sided Kruskal-Wallis test P < 0.0001, α: P < 0.0001, n = 50, 36, 33 FAs for each condition, respectively. k tFAK integrated intensity (mean ± SD) at FAs on 5 kPa mPADs. ANOVA P < 0.0001, α: P = 0.0032, β: P = 0.0195, n = 49, 37, 33 FAs for each condition, respectively. l pY397-FAK integrated intensity (mean ± SD) at individual FAs on 5 kPa mPADs. Two-sided Kruskal-Wallis test P < 0.0001, α: P = 0.0002, β: P < 0.0001, n = 50, 36, 33 FAs for each condition, respectively. m Force (mean ± SD) at individual FAs on 14 kPa mPADs. Two-sided Kruskal-Wallis test P < 0.0001, α: P < 0.0001, n = 66, 48, 28 FAs for each condition, respectively. n tFAK integrated intensity (mean ± SD) at FAs on 14 kPa mPADs. ANOVA P < 0.0001, α: P < 0.0001, β: P = 0.0004, n = 66, 48, 33 FAs for each condition, respectively. o, pY397-FAK integrated intensity (mean ± SD) at individual FAs on 14 kPa mPADs. Two-sided Kruskal-Wallis test P < 0.0001, α: P < 0.0001, β: P < 0.0146, n = 66, 46, 28 FAs for each condition, respectively. p Force vs. tFAK intensity at individual FAs on 5 kPa. Linear regression: control P = 0.1230, n = 50 FAs; Y-27632 P = 0.1920, n = 37 FAs; PF-228 P = 0.9869, n = 38 FAs. q, Force vs. pY397-FAK intensity at individual FAs on 5 kPa. Linear regression: control P = 0.1072, n = 50 FAs; Y-27632 P = 0.2937, n = 36 FAs; PF-228 P = 0.8874, n = 33 FAs. r Force vs. tFAK intensity at individual FAs on 14 kPa. Linear regression: control P < 0.0001, n = 66 FAs, tFAK intensity = 194.6 × force + 1178; Y-27632 P = 0.8896, n = 48 FAs; PF-228 P = 0.2084, n = 28 FAs. s Force vs. pY397-FAK intensity at individual FAs on 14 kPa. Linear regression: control P < 0.0001, n = 66 FAs, pY397-FAK intensity = 219.7 × force + 2027; Y-27632 P = 0.7986, n = 46 FAs; PF-228 P = 0.2260, n = 26 FAs.

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