Figure 6

The 11A9 antibody may cross-react with SWI/SNF component SMARCC1/BAF155. (A) Stoichiometry analysis in HEK293T with iBAQ values generated with Maxquant normalized with the IgG isotype control. AGO2 is the bait and set at 1 as we used anti-AGO2 (11A9) for the IP. Error bars represent standard deviations of 3 individual IPs. (B) Label-free quantification of BAF155 A301-019A associated proteins compared to IgG control. Experiment was performed in triplicate in HEK293T cells. For the analysis a two sample T-test was applied. (C) Table with average Maxquant derived label free quantification intensities of 3 IP-MS experiments for BAF155 A301-019A and IgG with HEK293T wild type lysates. Mean LFQ values are times 10 billion and signals from AGO2 knock-out come from imputation. CV represents coefficient of variation. (D) Venn diagram representing the number of quantified and significant proteins in the SMARCC1 IP-MS and the AGO2 11A9 IP-MS and proteins that were quantified and significant in both experiments. (E) Stoichiometry analysis as in A with SMARCC1 as bait. SMARCC1 was set at 1 as we used anti-BAF155 (A301-019A) for the IP. Error bars represent standard deviations of 3 individual IPs. (F) Reciprocal co-IPs with AGO2 11A9 and BAF155 antibodies with anti-IgG as negative control in Wildtype and AGO2 knock-out HEK293T cells. All samples were loaded on the same gel and transferred to the same blot on which both the staining’s were performed with different detection methods. Uncropped images are shown in the supplementary.