Anti cdk6 3136s

Cell protein lysates were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred to 0.22 μm polyvinylidene fluoride (PVDF) membranes (Millipore, USA), and incubated with specific antibodies. Autoradiograms were quantified by densitometry (Quantity One software; Bio-Rad, CA, USA). GAPDH antibody was used as a control. Anti-E2F1 (ab179445, 1:1000), -CDK4 (ab199728, 1:1000), -CDK6 (ab199728, 1:1000), -Ki67 (ab16667, 1:1000), -Bcl-2 (ab182858, 1:1000), -ILF3 (ab131004, 1:1000), -MAP2K6 (ab33866, 1:1000), and -GAPDH (ab9485, 1:2000) antibodies were purchased from Abcam. Anti -CDK6 (3136s, 1:1000), -P21 (2947s, 1:1000) were purchased from Cell Signaling Technology.

Human GC cell lines (AGS, BGC-803, BGC-823, MKN-45 and SGC-7901)
were purchased from the Type Culture Collection of the Chinese Academy of
Sciences (Shanghai, China). The human normal gastric mucosal cell line GES-1 was
acquired from Biowit Technologies Corporation (Shenzhen, China). All cell lines
were cultured as described previously [14 (link)].
Mouse anti-cyclin B1(sc-245) and anti-Cyclin D1 (sc-8396) were
purchased from Santa Cruz (Santa Cruz, CA, USA), whereas mouse
anti-CDK4(#2906S)and anti-CDK6(#3136S)were purchased from Cell Signaling
Technology (CST, MA, USA). Rabbit anti-USP22 (55110–1-AP), anti-OCT1 (POU2F1)
(10387–1-AP), anti-E-cadherin (20874–1-AP), anti-γ-catenin (27872–1-AP),
anti-Vimentin (Ag0489), and anti-MMP2 (10373–2-AP) were purchased from
Proteintech (Wuhan, China). Rabbit anti-ERK1/2 (137F5), anti-p-ERK1/2
(Thr202/Tyr204), anti-AKT (C-terminal), and anti-p-AKT (Ser473) were purchased
from CST (MA, USA). Rabbit anti-GAPDH and anti-Ki-67 (EPR3610) were purchased
from Abcam (Abcam, Cambridge, UK).