Anti cyclinb1 antibody

Male C57BL/6 (8 to 10 wk old) mice used in the experiments were purchased from the Shanghai Lab Animal/Research Center (Shanghai, China). ConA was obtained from Sigma (MO, USA), and CCl4 was obtained from Sinopharm Chemical Reagent Co. Ltd (Shanghai, China). IL-22-FP was provided by GENERON Corporation Ltd. (Shanghai, China); recombinant human interleukin 22 (rh-IL-22) was obtained from Sino Biological Inc. (Beijing, China). Anti-CyclinB1 antibody was purchased from Abcam (Shanghai, China). Other antibodies used in this article, including anti-STAT3, anti-phospho-STAT3 (Tyr⁷⁰⁵), and proliferating cell nuclear antigen (PCNA) antibodies, were purchased from Cell Signaling Technology (Beverly, MA, USA).

Anti‐Arf1 antibody (#10790‐1‐AP and #20226‐1‐AP), anti‐Rab35 antibody (#11329‐2‐AP), anti‐HDAC6 antibody (#16167‐1‐AP), anti‐NAT10 antibody (#13365‐1‐AP), anti‐Ran antibody (#10469‐1‐AP), anti‐TPX2 antibody (#11741‐1‐AP), anti‐Myt1 antibody (#67806‐1‐lg), anti‐GAPDH antibody (#60004‐1‐lg), and anti‐α‐tubulin antibody (#11224‐1‐AP) were purchased from Proteintech (Rosemont, IL, USA). Anti‐GM130 antibody (#DF7556) was purchased from Affinity Biosciences (Cincinnati, OH, USA). Anti‐Aurora A antibody (#AF1708) was purchased from Beyotime (Shanghai, China). Anti‐Plk1 antibody (#37‐7000) was purchased from Invitrogen (Carlsbad, CA, USA). Anti‐Ac‐tubulin antibody (#T7451), anti‐α‐tubulin‐FITC antibody (#F2168), and Hoechst 33 342 were purchased from Sigma–Aldrich (St. Louis, MO, USA). Anti‐Myc tag (#ab18185), Alexa Fluor 594 Goat Anti‐Rabbit IgG (H + L) (#ab150080), Alexa Fluor 594 Goat Anti‐Mouse IgG (H + L) (#ab150116), anti‐CDK1 antibody (#ab18), anti‐cyclin B1 antibody (#ab181593), and Anti‐GM130 antibody (#ab52649) were purchased from Abcam (Cambridge, UK). Anti‐phospho‐Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) antibody (#2914T) and antiactin antibody (#3700) were purchased from Cell Signaling Technology (Danvers, MA, USA). Horseradish peroxidase–conjugated goat antirabbit/mouse antibodies (CW0103/CW0102) were purchased from CWBIO (Beijing, China).

Cells were washed in PBS and then collected in RIPA lysis buffer (Beyotime Biotechnology, China) for protein quantification followed by addition of sample buffer and heat denaturation at 95°C for 10 min. Protein lysates were separated using 10% SDS-polyacrylamide gel electrophoresis (PAGE) gels and transferred to Immobilon-P transfer membranes (Millipore, USA). Membranes was blocked with 1% BSA supplement with 0.05% Tween-20, followed by incubation with primary antibodies overnight at 4°C. After washing with TBST, PVDF membranes were incubated with HRP-conjugated antibody for 1 h at 37°C. Immune complexes were visualized using the enhanced chemiluminescence ECL kit (Merck Millipore, USA), and detected using ChemiScope3400 Mini (Clinx Science Instruments, China). The following primary antibodies were used: anti-CDC27 antibody (Abcam, USA), anti-cyclinA antibody (Abcam, USA), anti-cyclinB1 antibody (Abcam, USA), anti-cyclin B2 antibody (Abcam, USA), anti-cyclin D1 antibody (Abcam, USA), anti-securin antibody (Abcam, USA), anti-E-Cadherin antibody (Abcam, USA), anti-beta Catenin antibody (Abcam, USA), anti-beta Catenin (phospho T41 + S45) antibody (Abcam, USA), anti-vinculin antibody (Sigma, USA), anti-FAK antibody (RD, USA), anti-FAK (phospho S732) antibody (Abcam, USA), and anti-GAPDH (1:1000, Boster Biotechnology, China).