Fig. 5

Active integrin β3 distribution and colocalization with pFAK³⁹⁷ within focal adhesions are influenced by ABCA1 activity. A and C Representative confocal images of scrambled and ABCA1 KO cells (A) or DMSO- and probucol-treated cells (C) stained with an antibody recognizing pFAK³⁹⁷ followed by a secondary antibody conjugated with Alexa Fluor 488 (green) and antibody against active integrin β3 followed by secondary antibody conjugated with Alexa Fluor 647 (red). Scale bars represent 20 µm. B and D Magnified view of scrambled (ROI1) and ABCA1 KO (ROI2) cells (B) from (A) or of DMSO- (ROI3) and probucol-treated (ROI4) cells (D) from (C). White arrowheads point at pFAK³⁹⁷-rich active integrin β3 clusters. Scale bars represent 20 µm. E, F Quantification of the percentage of the cells with active integrin β3 clusters enriched in pFAK³⁹⁷ of scrambled and ABCA1 KO cells (E) or DMSO- and probucol-treated cells (F) (n = 30 from three independent experiments, unpaired Student t-test). G, H Quantification of clusters number of active integrin β3 of scrambled and ABCA1 KO cells (G) from (A) or DMSO- and probucol-treated cells (H) from (C) (n = 30 from three independent experiments, unpaired Student t-test). I, J Quantification of the clusters size of active integrin β3 of scrambled and ABCA1 KO cells (I) from (A) or DMSO- and probucol-treated cells (J) from (C) (n = 30 from three independent experiments, unpaired Student t-test). ROI region of interest. Data are mean ± SD. **p ≤ 0.01 and ****p ≤ 0.0001