Figure 7. Expression of SS18-SSX leads to CBAF complex reductions and relative overabundance of PBAFs and GBAFs.

A. Quantitative LICOR WBs of BAF components in nuclear extracts of EXPI293 cells transfected with either SS18-SSX or SS18 (n = 5 each) presenting the log transformed 2-tailed Student’s t-test p-value of the difference between and the ratio of protein in the fusion-transfected versus SS18-transfected cells.

B. WBs of glycerol gradient fractions of EXPI293T cells transfected with SS18-SSX or SS18.

C. LICOR quantitative WBs from whole cell lysates (WCL) collected from HSSYII human SS cells subjected to 7 days of shRNAs (2 sequences each, n = 5 for each sequence of each shRNA) directed against control (Renilla, shRen) or the fusion (SS18-SSX, shSSX), with BAF subunits color-coded by BAF, presented as log-transformed paired 2-tailed t-test p-values and ratios of fusion knock-down over control knock-down.

D. LICOR quantitative WBs of WCLs from SYO-1 human SS cells after knock-down of the fusion or control for 7 days.

E. Fractional abundances of BAF subtypes defined by optical densitometry-quantified gradients of SMARCC1 (as in 6C) for HSSYII cells subjected to shRNAs against the fusion or control. (p-values from 2-tailed paired t-tests, n = 4 for each shRNA).

F. WBs of nuclear extract (NE) with the paired chromatin fraction (CF, protein that stays with the insoluble chromatin pellet after NE) of proteins after 7 days of fusion or control knock-down.

G. LICOR quantitative WB abundances presented as paired t-test p-values and ratios of fusion over control knock-down in each of the NE and CF components of HSSYII cells after 7 days.

H. LICOR quantitative WB-defined proteins in WCLs presented as the ratios of MG132-treated over DMSO vehicle-treated (veh) cells after week long shRNA depletion of SS18-SSX or control Renilla. (n = 5 for each condition, n = 10 for each group, 2-tailed heteroscedastic t-test comparing the ratios for each protein by knock-down group)

I. LICOR quantitative WB-defined proteins in WCLs presented as of the ratios of MG132-treated over DMSO vehicle-treated (veh) cells from SS (or SS18-SSX-transfected) and control (or SS18-transfected) cell lines (n = 5 each, p-values are from 2-tailed Student’s t-tests comparing each to HEK293T untransfected control cells).

J. Model schematic of the impact of SS18-SSX expression on BAF componentry and relative abundance of BAF subtypes.