Anti lamin b ab16048

The antibodies used were: anti-phospho JNK (Thr183/Tyr185) (#4668), anti-phospho STAT3 (Tyr705) (#9131), anti-STAT3 (#8719), anti-phospho p38 MAPK (Thr180/Tyr182) (#9211), anti-p38 MAPK (#9212), anti-phospho Foxo1 (#9461) and anti-Akt (#9272) from Cell Signaling Technology (MA, USA); anti-phospho IGFIR (Tyr1165/1166) (sc-101704), anti-JNK (sc-571), anti-phospho-Akt1/2/3 (Ser473) (sc-7985-R), anti-caspase 1 (sc-514), anti-Nrf2 (sc-722) and anti-Keap1 (sc-33569) from Santa Cruz (Palo Alto, CA); anti-phospho IRS1 (Tyr1179) (07-844), anti-phospho IRS1 (Ser 307) (07-247), anti-IRS1 (06-248), anti-p85α (06-195) and anti-HO1 (AB1284) antibodies from Merck Millipore (Merck KGaA, Darmstadt, Germany); anti-β-actin (A-5441) antibody from Sigma Chemical Co. (St Louis, MO); anti-Lamin B (aB16048) and FasL (aB68338) from Abcam (Abcam, Cambridge, UK). Anti-IGFIR antibody was a gift of S. Pons (CSIC, Spain).

Forty micrograms of nuclear and cytosolic extracts from INS-1 832/13 cells were subjected to 10% discontinuous SDS-PAGE before transferring onto PVDF membrane (PALL). The blots were incubated with appropriate dilution of anti-Sp1 (sc-59), anti-USF1 (sc-22), anti-USF2 (sc-862), anti-ChREBP (sc-21189) [Santa Cruz Biotechnology], anti-phospho Sp1 (ab59257), anti-tubulin (ab6046) or anti-laminB (ab16048) [Abcam], in 5% BSA in TBS-T overnight. The blots were washed with TBS-T before incubating with appropriate secondary antibodies conjugated with HRP for 3 h. The immunoreactive bands were visualized by an enhanced chemiluminescence using ECL Western substrate kit (Pierce).