Extended Data Fig. 6: Long-term effect of treatment of TEX cells with RB-3.
From: Small-molecule inhibitors targeting Polycomb repressive complex 1 RING domain
a, Western blot detection of H2Aub in TEX cells treated with increasing doses of RB-3 and RB-nc for 21 days. Representative blot out of two replicates. b, Western blot detection of total cellular ubiquitination levels using ubiquitin specific antibody in TEX cells treated with RB-3 for 21 days. Representative blot out of three replicates. c, Flow cytometry analysis of CD34 in TEX cells treated with RB-3 for 7 days. On day 7, cells were stained with APC/CY7-conjugated human anti-CD34 antibody and analyzed by FACS. Representative histograms of two independent experiments. d, Flow cytometry analysis of CD34 and CD38 in TEX cells treated with RB-3 (top panel) and RB-nc (bottom panel) for 21 days. On day 21, cells were stained with APC/CY7-conjugated human CD34 and PE-conjugated CD38 antibodies and analyzed by FACS. Representative histograms of two independent experiments. e,f, Flow cytometry analysis of myeloid differentiation marker CD11b/ITGAM in TEX cells treated with RB-3 for 7 days (e) and with RB-3 (f, top panel) and RB-nc (f, bottom panel) for 21 days. Cells were stained with Pacific Blue human CD11b antibody and analyzed by FACS. Representative histograms of two independent experiments. g, Flow cytometry analysis of dendritic cells differentiation marker CD86/B7-2 in TEX cells treated with RB-3 (top panel) and RB-nc (bottom panel) for 21 days. On day 21, cells were stained with Super Bight 436 conjugated human CD86 antibody and analyzed by FACS. Representative histograms of two independent experiments.
