Ag490 s1143

All the cells were incubated at 37 °C with 5% CO₂. GBM8401, GB8901, and DBTRG-05MG cells were cultured in RPMI medium plus 10% fetal bovine serum (FBS). The U87-MG and SVGp12 cells were cultured in modified Eagle’s medium (MEM) plus 10% FBS. G5T cells were cultured in Dulbecco’s MEM medium plus 10% FBS. GBM8401, GBM8901, U87-MG, G5T, and DBTRG-05MG were GBM cell lines, and SVGp12 was a glial cell. The cells were treated with AG490 (S1143, Selleckchem, Houston, TX, USA) at 0, 100, 200, 300, 400, and 500 nM concentrations.

The reagents used in this study were obtained from the indicated suppliers: IFN-γ, IL-1β, TNF-α, IL-4, IL-13, and TGF-β from R&D Systems (Minneapolis, MN, USA); antibodies against α-SMA (ab7817) from Abcam (Cambridge, UK); antibodies against GAPDH (sc47724) and TSG-6 (sc377277) from Santa Cruz Biotechnology (Santa Cruz, CA, USA); antibodies against STAT1 (9172S), pSTAT1 (9167S), STAT3 (9139S), pSTAT3 (9145S), and Smad2/3 (8685S) from Cell Signaling Technology (Danvers, MA, USA); antibodies against pSmad2/3 (PA5-110155) from ThermoFisher Scientific (Waltham, MA, USA); chemical inhibitors for JAK1 (Abrocitinib, HY-107429; MedChemExpress, Princeton, NJ, USA) and JAK2 (AG-490, S1143; Selleck Chemicals, Houston, TX, USA); scramble and TSG-6 siRNA (Santa Cruz). All other materials were purchased from Sigma-Aldrich (St. Louis, MO, USA) unless otherwise indicated.

The human colon cancer cell lines HCT116 and LoVo were obtained from ATCC (Manassas, VA, USA). HCT116 and LoVo cells stably expressing PRL-3 and control cells (NC) were previously established [40 (link)]. NF-κB inhibitor Bay 11–7082 (S2913) and Jak2 inhibitor AG490 (S1143) were obtained from Selleckchem (Houston, TX, USA). IL-6 (TP723240) was obtained from OriGene Technologies (Rockville, MD, USA). Recombinant human IL-1RN (ag1277) and anti-GAPDH (10494-1-AP) were from Proteintech Group (Chicago, IL, USA). Anti-Myc tag (AB103) was from TianGen Biotech (Beijing, China). Mouse anti-PRL-3 monoclonal antibody was generated by immunizing mice with KLH-conjugated full-length human PRL-3. Anti-phosphoserine/threonine (612548) was from BD Bioscience (San Jose, CA, USA). Anti-p65 (sc-372) was from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-phosphotyrosine (9411), anti-pS536-p65 (3033), anti-stat3 (9139), anti-pS705-stat3 (9145), anti-pS727-stat3 (9134), anti-Jak2 (3230), anti-pY1007/1008-Jak2 (3776), anti-Erk (9102), anti-pT202/Y204-Erk1/2 (9106), anti-EGFR (4267), anti-pY1045-EGFR (2237), anti-pY1068-EGFR (3777), anti-pY527-Src (2105), anti-pT180/Y182-p38 (4511), anti-pT581-MSK1 (9595), and anti-pT334-MAPKAPK2 (3007) were purchased from Cell signaling (Danvers, MA, USA).