(A) Outline of method for quantifying the number of molecules of a specific protein per cell from different cell lines.
(B) Quantifications of number of ACTL6A molecules per cell compared to SMARCA4/SMARCA2. SCC cell lines: FaDu (head-and-neck), NCI-H520 (lung), and T.T (esophageal). KC: primary normal human keratinocytes. n=3 experiments. Error bars indicate SEM. *P < 0.05.
(C) Co-immunoprecipitation (Co-IP) experiments using SMARCA4 antibody. Shown are Western blots and quantifications of relative levels of BAF subunits co-IP’d by SMARCA4 in SCC (FaDu) cells versus primary human keratinocytes (KC). n=3 experiments. Error bars indicate SEM. *P < 0.05. n.s.: not significant.
(D) Co-IP experiments using SMARCA4 antibody in primary human keratinocytes (KC) transduced by lentivirus for ACTL6A overexpression and vector control. Shown are Western blots and quantifications of relative levels of co-IP’d BAF subunits normalized to vector control. n=3 experiments. Error bars indicate SEM. *P < 0.05. n.s.: not significant.
(E) Quantifications for co-IP experiments by SMARCA4 antibody. Relative levels of co-IP’d ACTL6A in ACTL6A-overexpressing condition normalized to vector control. FaDu: SCC cell line. KC: primary human keratinocytes. HEK293T: human embryonic kidney 293T cells. Error bars indicate SEM. *P < 0.05. n=2–3 experiments.