Anti erk

The GFP-positive embryos were raised to adults and out-crossed to wild-type fish for testing germline transmission. Transgenic fish were dissected to expose the abdominal area and liver morphology observed under the SMZ1600 stereomicroscope (Nikon, Tokyo, Japan). Liver tumor in transgenic fish was defined as GFP-marked liver which was enlarged to at least twice the size of a WT normal liver. Liver samples were then fixed in 4% paraformaldehyde (PFA) for at least 24 hours by dehydration through a series of graded ethanol solutions and embedding in paraffin. Four µm- thick sections were cut and stained with hematoxylin and eosin (H&E) for histological analysis. The following antibodies were used for immunohistochemical study: anti-CK19 (Abcam, Cambridge, UK, ab9221), anti-PCNA (Servicebio, Wuhan, China, GB13010–1), anti-MEK1 (HuaAn, Hangzhou, China, ET1603-20), anti-MEK2 (HuaAn, Hangzhou, China, ET1612-6), anti-ERK (Servicebio, Wuhan, China, GB13003-1), anti-pMEK1/2 (Cell Signaling, Danvers, MA, #9154), anti-pERK (Servicebio, Wuhan, China, GB13004-1).

Whole-cell protein were isolated from Primed H9 and Naïve-like H9. Protein samples were incubated with the following primary antibodies in 5%BSA: anti-ERK (Servicebio, Wuhan, China, GB13003–1), anti-JNK (Epitomics, 3496-s), anti-P38 (ABCAM, ab31828), and anti-β-actin (Servicebio, Wuhan, China, GB13001–1). Secondary antibodies were HRP-linked goat anti-mouse, goat anti-rabbit (Servicebio, Wuhan, China, GB23303). Blots were developed using ECL (Servicebio, Wuhan, China, G2014). The primary antibodies used in our study were listed in Additional file 2: Table S1.