Fig. 4
Stable recruitment of active form of FAK in FAs is dependent on 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 secondary antibody conjugated with Alexa Fluor 488 (green) and antibody against total actin followed by secondary antibody conjugated with Alexa Fluor 647 (red) after fixation. 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 enriched in pFAK397 FAs. Scale bars represent 20 µm. E, F Quantification of cells number exhibiting enriched pFAK397 FAs in scrambled and ABCA1 KO cells (E) or DMSO- and probucol-treated cells (F) expressed as % of cells with enriched pFAK397 FAs from the total number of cells analyzed (n = 30 from three independent experiments, unpaired Student t-test). G and I Representative immunoblots of pFAK397 and total FAK levels in scrambled and ABCA1 KO cells (G) or DMSO- and probucol-treated cells (I). Vinculin was used as a loading control. H and J Quantification of pFAK397 relative levels of scrambled and ABCA1 KO cells (H) from (G) or of DMSO- and probucol-treated cells (J) from (I). (n = 3 independent experiments, unpaired Student t-test). ROI region of interest. Data are mean ± SD. ns non-significant and ****p ≤ 0.0001