Supplementary Figure 6: Secondary siRNA validation and effects of PI4,5P2 modulation.

(a) Single cell quantification (n = 8070 cells analysed, averaging 224 + /- 63 (stdev) per condition) of reticular to focal adhesion integrin β5 ratios for 4 independent siRNAs targeting controls or PIP regulators. Boxplot centre and box edges indicate median and 25^th or 75^th percentiles, respectively, while whiskers indicate the median + /- 1.5*IQR (inter-quartile range) or the most extreme observations within these limits. Boxplot notches approximate 95% confidence intervals (see methods for details). P-values reflect two-sided unpaired Mann Whitney U testing with Holm-Bonferroni correction from multiple tests. Data derived from 2 biologically independent experiments. Source data is available in Supplementary Table 1 (b) Representative images of U2OS cells stained with DAPI (nuclei) and deep red cell mask membrane dye and showing integrin β5-2GFP and mCherry-vinculin fluorescence following treatment with DMSO, 10 mM Neomycin or 25 μM LY294002. Images in B representative of n = 3 biologically independent experiments. Scale bars = 50 µm