Sc 7866

M-MØ and GM-MØ cell lysates were subjected to SDS-PAGE (30-50 μg unless indicated otherwise) and transferred onto an Immobilon-P polyvinylidene difluoride membrane (PVDF; Millipore). After blocking the unoccupied sites with 5% non-fat milk diluted in Tris-Buffered Saline plus Tween 20 (TBS-T), protein detection was carried out with antibodies against LXRα (PPZ0412; Biotechne), LXRβ (PPK8917; Biotechne), MAFB (HPA005653, Sigma Aldrich) or c-MAF (sc-7866; Santa Cruz Biotechnology). Protein loading was normalized using an antibody against GAPDH (sc-32233; Santa Cruz Biotechnology) or vinculin (V9131; Sigma-Aldrich). Quimioluminiscence was detected in a Chemidoc Imaging system (BioRad) using SuperSignal™ West Femto (ThermoFisher Scientific).

The HEK-293T cells (2 × 10⁶ cells/well) were transected with RORγ-Flag (5 μg) and or Blimp-1-Myc (5 μg) plasmids. According to the standard procedure, immunoprecipitation and immunoblotting were performed to analyze the combination of c-Maf (1:500, sc-7866, Santa Cruz, Dallas, TX, USA), Blimp-1 (1:500, sc-47732, Santa Cruz) and RORγ (1:500 sc-81371, Santa Cruz).

Single-cell suspensions of spleen, lymph nodes and thymus from Cic^f/f and Cic^f/fVav1-Cre mice were used as total immune cells in lymphoid organs. Naïve CD4⁺ T cells were activated by anti-CD3 and anti-CD28 and collected on days 0 (unstimulated), 1, 2 and 3. B220⁺ B and CD4⁺ T cells from Cic^f/f and Cic^f/fCd4-Cre mice were prepared using MACS (130-049-501 and 130-104-454, Miltenyi Biotec) according to the manufacturer’s protocol. Naïve CD4⁺ T, effector/memory CD4⁺ T, T_FH and non-T_FH cells were purified from total CD4⁺ T cells by FACS sorting using the MoFlo-XDP. Splenic CD19⁺ B and CD3⁺ T cells were prepared from Cic^f/f and Cic^f/fCd19-Cre mice by FACS sorting using the MoFlo-XDP. Total lysates from 0.2 to 1.0 × 10⁶ cells of each cell population were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS–PAGE). The primary antibodies used are as follows. anti-CIC (1:500 diluted, homemade)28 (link), anti-ETV4 (1:1,000 diluted, 10684-1-AP, ProteinTech), anti-ERM/ETV5 (1:1,000 diluted, ab102010, Abcam), anti-MAF (1:750 diluted, sc-7866, Santa Cruz Biotechnology), anti-β-actin (1:2,000 diluted, sc-47778, Santa Cruz Biotechnology) and anti-GAPDH (1:2,000 diluted, sc-32233, Santa Cruz Biotechnology). Raw western blot images are presented in Supplementary Figs 18 and 19.