Klf5 21017 1 ap

DCC-2036 (Rebastinib) was purchased from Selleck (Houston, TX). The compound was dissolved in DMSO at a stock concentration of 20 mM and stored at −20 °C. DCC-2036 (0 μM) means DMSO treatment with equivalent volume. Recombinant Human GAS6 (C-Fc) #C12W was purchased from Novoprotein (Suzhou, Jiangsu, China). Recombinant Mouse GAS6 Protein (His Tag) #58026-M08H was purchased from Sino Biological (Shanghai, China). Cycloheximide and MG132 were purchased from Sigma-Aldrich (St Louis, MO, USA). The following antibodies were used for immunoblotting: phospho-AXL (Y702) #5724, phospho-AKT (S473) #4060, AKT #4691,phospho-GSK3β (S9) #5558, GSK3β #12456, β-actin #4970, purchased from Cell Signaling Technology (Beverley, MA); ALDH1A1 #15910-1-AP, OCT4 #11263-1-AP, KLF4 #11880-1-AP, SOX2 #11064-1-AP, KLF5 #21017-1-AP, NANOG #14295-1-AP, AXL #13196-1-AP, MET #25869-1-AP, FGF-BP1 #25006-1-AP purchased from Proteintech (Wuhan, Hubei, China); NF-κB (P65) #ab16502, purchased from Abcam; FGF-BP1 #AF1593 purchased from R&D; phospho-KLF5 (S303) #AF7042, phospho-AXL (Tyr702) # AF8523 purchased from Affinity (Changzhou, Jiangsu, China). APC anti-mouse CD24 #138506 purchased from Biolegend (San Diego, CA, USA), PE Rat anti-mouse CD44 #553134, PE Mouse Anti-Human CD24 #555428, APC Mouse Anti-Human CD44 #559942 purchased from BD Pharmingen (Franklin Lakes, New Jersey, USA).

To verify the expression of GOIs, proteins were extracted by lysed cells using RIPA Lysis and Extraction Buffer (#89900; ThermoFisher) and the concentrations of protein samples were quantified with the Pierce BOA Protein Assay Kit (#23225; ThermoFisher). The western blot analysis was used to determine the expression of protein-of-interest. The intensity of protein bands was detected by Immobilon Western Chemiluminescent HRP Substrate (#WBKLS0500; MilliporeSigma, Burlington, MA) using the ChemiDoc Imaging System. The antibody targeting b-actin (8H10D10, #3700) was purchased from the Cell Signaling Technology (Danvers, MA), human ACE2 (AF933) was purchased from R&D Systems, DAZAP2 (G-4, sc-515182) was purchased from Santa Cruz Biotechnology (Dallas, TX), KLF5 (21017-1-AP) was purchased from Proteintech Group (Rosemont, IL) and the monoclonal ANTI-FLAG antibody (M2, #F3165) was purchased from MilliporeSigma.