P stat3 9131

Antibodies to pan-RAS (ab52939), H-RAS (ab97488), N-RAS (ab77392), K-RAS (ab137739), Raf1 (ab137435), p-JAK1 (sc-101716), p-ERK1/2 (sc-7383), AKT (sc-5298), STAT3 (sc-482), JAK1 (sc-7228), SRC (sc-8056), OCT4 (sc-9081), SOX2 (ab97959) and β-actin (sc-47778), anti-mouse IgG-HRP, anti-goat IgG-HRP, and anti-rabbit Ig-HRP were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies to Anti-Goat Alexa Fluor 488, and anti-mouse Alexa Fluor 546 were purchased from Invitrogen (Carlsbad, CA, USA). Antibodies to ERK (4695), CD44 (ab157107), Nanog (ab21624), Slug (ab27568), Zeb1 (ab124512), Vimentin (ab8978), and Fibronectin (ab6329) were purchased from Abcam (Cambridge, UK). Antibodies MEK1/2 (8727), p-MEK1/2 (9121), p-JNK (9251), JNK (9252), p-Src (9211S), p-AKT (4060), Snail (3879S), P38 (9212), p-P38 (9211S) and p-STAT3 (9131) were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibodies to GPR110 were purchased from LSbio. Antibodies to Active-RAS were purchased from NewEast, Antibodies to p-Raf1 were purchased from MyBioSource. Antibodies to Ki-67 were purchased from Actis. Antibodies to N-cadherin and β-catenin were purchased from BD biosciences. Vector of GPR110 were purchased from origene (ADGRF1, NM_153840, RC222706).

Polybrene and Puromycin were purchased from Sigma. The primary antibodies used in this study: SNF5(20,654-1-AP) and GAPDH (10,494-1-AP) were purchased from Proteintech. PD-L1(13,684), p21(2947), cyclin D1(55,506), STAT3(9139) and p-STAT3(9131) were purchased from Cell Signaling Technology. PD-L2 (ab187662) and IDO1(ab211017) were purchased from Abcam.

The sciatic nerve of C57BL/6 mice is composed from elements of L3 and L4 segments of spinal cord with a smaller contribution from L5 segment [14] (link). Therefore, after euthanizing each mouse with pentobarbital, the L3-L5 section of its spinal cord was quickly removed and homogenized in ice-cold RIPA lysis buffer. After concentration determination, protein samples were separated in 10% SDS-PAGE and electrotransferred onto PVDF membranes. Protein samples were then probed with antibodies for extracellular signal-regulated kinase (ERK, 4695, Cell Signaling), phosphorylated ERK (pERK, 4511, Cell Signaling), c-Jun N-terminal kinase (JNK, 9258, Cell Signaling), phosphorylated JNK (pJNK, 4668, Cell Signaling), p38 (9212, Cell Signaling), phosphorylated p38 (p-p38, 4511, Cell Signaling), AKT (9272, Cell Signaling), phosphorylated AKT (pAKT, 4060, Cell Signaling), Signal transducer and activator of transcription 3 (STAT3, 9132, Cell Signaling), phosphorylated STAT3 (pSTAT3, 9131, Cell Signaling), or p65 (4764, Cell Signaling) with alpha-tubulin (T5168, Sigma) used as a loading control. The membranes were visualized with horseradish peroxidase (HRP)-conjugated secondary antibodies (anti-rabbit IgG and anti-mouse IgG, Sigma).