Jnk 10023 1 ap

Proteins were separated on 10% or 12% SDS-PAGE gel and transferred to PVDF membranes (Millipore, IPVH00010), which were blocked with 5% BSA (Genview, FA016). Next PVDF membranes were incubated with the following antibodies: SREBP1 (PA1-337, ThermoFisher), Bax (50599-2-AP, Proteintech), Bcl2 (12789-1-AP, Proteintech), Caspase 3 (19677-1-AP, Proteintech), Caspase 9 (#9502, Cell Signaling Technology), AKT (#4691, Cell Signaling Technology), p-AKT (#4060, Cell Signaling Technology), JNK (10023-1-AP, Proteintech), p-JNK (80024-1-RR, Proteintech) and β-Actin (AC004, Abconal). After washed 3 times with TBST buffer, PVDF membranes were then incubated with horseradish peroxidase-conjugated anti-rabbit or anti-mouse secondary antibodies (Thermo Fisher Scientific). Western blot band were visualized by digital gel image analysis system (TANON 5500) and Pro-Light chemiluminescence detection kit (TIANGEN, PA112-01).

Antibodies for OTUD7B (16605-1-AP) and JNK (10023-1-AP) were from Proteintech Group, Inc. (Chicago, USA). Antibody for GAPDH (AC033) was from Abclonal (Wuhan, China). Antibodies for p70S6K (2708), p-p70S6K T389 (9206), p-p70S6K T421/S424 (9204), ERK1/2 (9102), p-ERK1/2 T202/Y204 (9101), p-Akt S473 (4060), p-p38 T180/Y182 (9215), and p-JNK T183/Y185 (4668) were purchased from Cell Signaling Technology (Boston, USA). Antibody for p38 (35478) was from SAB. Antibody for Akt1(SC-5298) was from Santa Cruz Biotechnology (Dallas, USA). Human VEGFA monoclonal antibody (M808) and Biotin-labeled VEGFA polyclonal antibody were purchased from Thermo Fisher Scientific (Waltham, USA). EGF was from Peprotech (Suzhou, China).