Anti xbp1s 24868 1 ap

To evaluate protein expression the following primary antibodies were used: rabbit polyclonal anti-phospho-eIF2α (Ser51) (9721; Cell Signaling, Danvers, MA, USA), rabbit polyclonal anti-eIF2α (9722; Cell Signaling, Danvers, MA, USA), rabbit polyclonal anti-XBP1s (24868-1-AP; Proteintech, Rosemont, IL, USA), anti-BiP/GRIP78 (11587-1-AP; Proteintech), rabbit polyclonal anti-Caspase 9 (10380-1-AP; Proteintech, Rosemont, IL, USA), rabbit polyclonal anti-c-Myc (10828-1-AP; Proteintech, Rosemont, IL, USA), mouse monoclonal anti-γ-H2AX (Ser 139) (sc-517348; Santa Cruz Biotechnology, Dallas, TX, USA), mouse monoclonal anti-BRCA-1 (OP92; EMD Millipore, Burlington, MA, USA), mouse monoclonal anti-RAD51 (G-9) (sc-377467; Santa Cruz Biotechnology, Dallas, TX, USA), and rabbit monoclonal anti-PARP (46D11) (9532; Cell Signaling, Danvers, MA, USA). Mouse monoclonal anti-β-actin (A5316; Sigma-Aldrich, Burlington, MA, USA) was used as loading control. The goat anti-Mouse IgGP Peroxidase Conjugate (401215; Sigma-Aldrich, Burlington, MA, USA) and the goat anti-Rabbit IgG Peroxidase Conjugate (DC03L; Sigma-Aldrich, Burlington, MA, USA) were used as secondary antibodies.

Cell or tissue lysates and prestained protein molecular weight markers (#26612, Pierce, USA) were separated by sodium dodecyl sulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE), followed by transferring onto polyvinylidene fluoride (PVDF) membranes. The membranes were blocked in Tris‐buffered saline with 0.5% Tween 20 (TBST) containing 5% bovine serum albumin (BSA) and probed with primary antibodies overnight at 4℃. Then, the membranes were washed three times and incubated with the horseradish peroxidase (HRP)‐conjugated secondary antibodies goat‐anti‐rabbit IgG (#7074) and horse‐anti‐mouse IgG (#7076). The primary antibodies used in the study were anti‐p300 (#86377), anti‐p‐IRE1α (#PA1‐16927, Invitrogen, USA), anti‐IRE1α (#3294), anti‐XBP1s (#24868‐1‐AP, Proteintech, USA), anti‐acetylated lysine (Ac‐K; #9681), anti‐activating transcription factor 4 (ATF4; #11815), anti‐glucose regulated protein 94 (GRP94; #2104), anti‐binding immunoglobulin protein (BIP; #3183), anti‐C/EBP homologous protein (CHOP; #2895), anti‐ubiquitin (#3933), anti‐Herpud1 (ab150424, Abcam, USA), and anti‐GAPDH (#2118). Antibodies whose supplier is not stated were purchased from Cell Signalling Technology (USA). Immunoreactivity was visualized by enhanced chemiluminescence (ECL, #34577, Pierce, USA).