Map3k11

Total protein was extracted from cells after transfection using lysis buffer for western and IP (KeyGen, KGP701, Nanjing, China), and protein concentration was measured using a BCA protein assay kit (Beyotime, P0010, Shanghai, China). Proteinsamples were electrophoresed using SDS-PAGE and then transferred for incubation at 4ºC overnight with the following specific primary antibodies: p53 (ImmunoWay, YT3528, TX, USA), Bax (ImmunoWay, YT0455, TX, USA), Bcl-2 (ImmunoWay, YM3041, TX, USA), active caspase-3 (Abcam, ab32042, Cambridge, UK), PTPN1 (Abcam, ab75856, Cambridge, UK), MAP3K11 (Abcam, ab51068, Cambridge, UK), JNK1/2/3 (Bimake, A5005, TX, USA), c-Jun (Bimake, A5730, TX, USA), and GAPDH (ImmunoWay, YM3445, TX, USA). The samples were then incubated with HRP-conjugated anti-rabbit IgG antibody (CST, #7074, MA, USA) at room temperature for 1 h. Finally, proteins were detected using ECL (KeyGEN, KGP902, Nanjing, China). GAPDH was used as an internal reference. Image J was used for quantification of all Western blot bands.

Tumor xenografts were fixed in paraformaldehyde (Servicebio, G1101, Wuhan, China) for 12 h and embedded in paraffin. After that, the KeyGEN One-Step IHC Assay (KeyGen, KGOS60-KGO300, Nanjing, China) was used to stain 5-μm-thick paraffin sections, which were then incubated with the following antibodies: Brdu (Servicebio, GB12051, Wuhan, China), Ki-67(ImmunoWay, YT2467, TX, USA), PTPN1(Abcam, ab75856, Cambridge, UK), MAP3K11(Abcam, ab51068, Cambridge, UK), JNK1/2/3 (Bimake, A5005, TX, USA), and c-Jun (Bimake, A5730, TX, USA). The paraffin sections were then photographed.