Cyclind1 2978s

WB was performed as previously described (23 (link)). Primary antibodies specific for Prp19 (ab126776, 1:1,000) and YAP (ab52771, 1:1,000) were purchased from Abcam; Primary antibodies specific for GAPDH (2118s, 1:2,000), cyclinD1 (2978s, 1:1,000), MMP9 (13667s, 1:1,000), E-cadherin (3195s, 1:1,000) were purchased from Cell Signaling Technology (Beverly, USA). A primary antibody specific for Actin was purchased from Yeasen (30101ES10, 1:5,000). Primary antibodies specific for CTGF (sc-101586, 1:200), FGF1 (sc-55520, 1:100) were purchased from Santa Cruz Biotechnology (USA). The results were quantitatively analyzed through Image J software (X64, v. 2.1.4).

Cells and tissues were collected and lysed with RIPA buffer containing phenylmethanesulfonylfluoride (Beyotime, China). Equivalent amounts of total protein were separated by 10% SDS-PAGE (Servicebio, China), then transferred to PVDF membranes (Servicebio, China). The membranes were then blocked in TBST containing 5% nonfat milk, and the membranes were incubated with primary antibodies overnight at 4 °C. The membranes were then incubated with secondary antibodies. An ECL kit (Multisciences, China) was used to detect proteins. Primary antibodies against FOXO1 (2880) and cyclin D1 (2978S) were purchased from Cell Signaling Technology. Antibodies against p21 (ab109520) and GAPDH (ab9485) were purchased from Abcam.

We purchased Akt and p-Akt (catalog nos. 4691S and 9271T), Bax and Bcl-2 (2772T and 2872T), mTOR and p-mTOR (2983S and 5536S), nuclear factor κB (NF-kB) p65 and 0-NF-kB (8242S and 3033S), Cox2 (12282S), 4EBP1 and p-4EBP1 (#9644 and #9451S), Cyclin D1 (2978S), CDK6 (13331T), and β-actin (A2228) as a primary from Cell Signaling Technology (Danvers, MA, USA) and anti-mouse and rabbit IgG (catalog no. A4416 and A0545) as a secondary antibody from Sigma Aldrich.