MSIE cells and intestinal epithelial cells isolated from mice were solubilized in cell lysis buffer to prepare total cellular lysates. Nuclear fractions were prepared using NE-PER Nuclear and Cytoplasmic Extraction Reagents (Thermo Scientific) according to the manufacturer’s instructions. The protein concentration was determined using a BCA protein assay kit (Thermo Fisher Scientific Inc.).
Cellular and nuclear lysates were mixed with Laemmli sample buffer and proteins were separated by SDS-PAGE for Western blot analysis using anti-phospho-Ser473(P)-Akt (Cell Signalling Danvers, MA), anti-phospho-Tyr1068-EGFR (Cell Signalling), anti-total EGFR (Millipore, Billerica, MA), anti-APRIL (Thermo scientific), anti-BAFF (Enzo Biochem, Inc, Farmingdale, NY), anti-AID (eBioscience), anti-p65 (Santa Cruze, Dallas, TX), anti-fibrillrin (Santa Cruze), and anti-β actin (Sigma-Aldrich) antibodies.
Cellular and nuclear lysates were mixed with Laemmli sample buffer and proteins were separated by SDS-PAGE for Western blot analysis using anti-phospho-Ser473(P)-Akt (Cell Signalling Danvers, MA), anti-phospho-Tyr1068-EGFR (Cell Signalling), anti-total EGFR (Millipore, Billerica, MA), anti-APRIL (Thermo scientific), anti-BAFF (Enzo Biochem, Inc, Farmingdale, NY), anti-AID (eBioscience), anti-p65 (Santa Cruze, Dallas, TX), anti-fibrillrin (Santa Cruze), and anti-β actin (Sigma-Aldrich) antibodies.