Baf170

Collected cells were counted at collection point and lysed in 1 × Leammli buffer. Samples were separated on SDS-PAGE gels and transferred onto PVDF membranes (0.45 μm pore dimension). Membranes were blocked in 5% non-fat dry milk (VWR) for 1 h at room temperature. Subsequently, membranes were incubated with primary antibodies overnight at 4 °C. The following primary antibodies were used according to the manufacturers’ instructions: anti-AMPK (#5832, Cell Signaling Technology), phosphorylated AMPK (p-AMPK) (#8208, Cell Signaling Technology), BAF155 (#11956, Cell Signaling Technology), BAF170 (#12760, Cell Signaling Technology), BRG1 (#3538, Cell Signaling Technology), BRM (#11966, Cell Signaling Technology), Ezh2 (#5246, Cell Signaling Technology), INI1 (#8545, Cell Signaling Technology), PD-1 (#86163, Cell Signaling Technology), PD-L1 (#13684, Cell Signaling Technology), SUZ12 (#3737, Cell Signaling Technology), IL-6 (#12153, Cell Signaling Technology), and IL-1β (#12703, Cell Signaling Technology). The membranes were washed three times with TBS-T (0.01%) and incubated with HRP-conjugated secondary antibody (antirabbit IgG, #1705046, Bio-Rad) for 1 h in room temperature. After washing three times with TBS-T, the signal was detected using a chemiluminescent substrate (Western Bright Quantum, Advansta) and visualized using a UVTech gel blot imaging system.

Rabbit antisera to Brg1 and MyoD were previously described (65 (link), 82 (link)). Pan-calcineurin A (catalog no. 2614), Baf170 (catalog no. 12769), Baf250A (catalog no. 12354), and TBP (catalog no. 8515) antibodies were from Cell Signaling Technologies (Danvers, MA). GAPDH antibody (catalog no. G9295) was from Sigma-Aldrich (St. Louis, MO). MF20 antibody was from the Developmental Studies Hybridoma Bank (Iowa City, IA). Brg1 antibody (G-7; catalog no. sc-17796) was from Santa Cruz Biotechnologies (Santa Cruz, CA) and was used for Western blotting and immunoprecipitation experiments.

Antibodies against FLAG (F1804, Sigma Aldrich), IPMK (custom rabbit polyclonal antibody, raised against a mouse IPMK peptide corresponding to amino acids 295–311 [SKAYSTHTKLYAKKHQS; Covance]; Kim et al., 2011 (link)), SMARCB1 (A301-087, Bethyl), BRG1 (ab110641, Abcam), BAF155 (11956, Cell Signaling Technology), BAF170 (12760, Cell Signaling Technology), BAF250A (12354, Cell Signaling Technology), BRM (11966, Cell Signaling Technology), PBAF/PBRM (A301-591A, Bethyl), a-TUBULIN (T5169, Sigma Aldrich), GST (2622, Cell Signaling Technology), LaminB1 (sc-365214, Santa Cruz Biotech), histone H3 (05–499, Sigma Aldrich), GAPDH (sc-32233, Santa Cruz Biotech), anti-DPF2 (ab128149, Abcam), anti-SMARCE1 (ab137081, Abcam), anti-SS18L1 (ab227535, Abcam), anti-ACTL6A (sc-137062, Santa Cruz Biotech), anti-SMARCD1 (sc-135843, Santa Cruz Biotech), anti-BCL7A (HPA019762, Atlas Antibodies), and anti-ACTB (TA811000, Origene) were used for immunoblotting. Antibodies against IPMK (homemade), SMARCB1 (A301-087, Bethyl), and rabbit IgG isotype control (02–6102, Invitrogen) and anti-FLAG M2 affinity gel (A2220, Sigma Aldrich) were used for immunoprecipitation, and GST (2622, Cell Signaling Technology) was used for pull-down assays. Antibodies against FLAG (F7425, Sigma Aldrich), BRG1 (ab110641, Abcam), and IgG (homemade) were used for CUT&RUN assays.