Anti senp1

Bortezomib was purchased from Janssen-Cilag (Buckinghamshire, UK) and was dissolved in dimethyl sulfoxide (DMSO). Anti-hnRNP K antibody was purchased from Cell Signaling Technology (New England Biolabs, Schwalbach, Germany) and Santa Cruz Biotechnology (Santa Cruz, Dallas, TX), anti-pan-SUMO, anti-SUMO-1, anti-SENP1, and anti-SENP2 antibodies were purchased from Abcam (Cambridge, MA), anti-c-Myc and anti-HA antibodies were purchased from Cell Signaling Technology, and anti-GAPDH antibody was purchased from GeneTex (Irvine, CA).

Western blotting analysis was carried out according to protocols as described previously.³⁸ In brief, the total brain protein extracts from the cerebral cortex of mice with lysis buffer were prepared for Western blotting. The equivalent amount of protein was separated by 10% acrylamide denaturing gels (SDS‐PAGE) and then transferred to PVDF membrane (Millipore). Membranes were blocked with fat‐free milk for 1 hour and incubated with primary antibodies as following: anti‐β‐Actin (1:5000, Sigma‐Aldrich); anti‐Calcineurin (made by oneself); anti‐spectrin (1:1000, Millipore); anti‐SENP1 (1:2000, Abcam); anti‐FADD (1:500, Santa Cruz); anti‐Fas‐L (1:500, Santa Cruz); anti‐bcl‐2 (1:500, Santa Cruz); anti‐ZO‐1 (1:1000, Invitrogen); and anti‐Occludin (1:1000, Invitrogen) at 4ºC overnight and then incubated with HRP‐conjugated secondary antibodies (1:5000, Life Science). The proteins were visualized by an enhanced chemiluminescence detection system (Amersham Life Science). The density of the bands was quantified with ImageJ software (NIH) and normalized to β‐Actin.