Dulbecco s modified eagle medium dmem hepes

Dulbecco’s Modified Eagle Medium (DMEM/HEPES, Cat #12430-054) and Penicillin/Streptomycin (Cat #15240062) were from Gibco (Carlsbad, CA). Fetal bovine serum (FBS) and G418 sulfate were obtained from Invitrogen (Carlsbad, CA). Rabbit antibody against β-actin (Cat #4970S) was from Cell Signaling (Beverly, MA). Rabbit antibody against NHE1 (Cat #ab67314) and rat antibody against CD8 (Cat #ab22378) were from Abcam Ltd. (Cambridge, MA). Rat anti-mouse CD31 (Cat #550274) was from BD Pharmingen (San Jose, CA). Rabbit anti-ionized calcium-binding adapter molecule 1 (iba1) was from Wako (Richmond, VA). PerCP/Cy5.5-CD45, PE-P2RY12, BV421-TGFβ, BV605-TNFα, APC/780-IL-1β, PE/Cy7-IL10, PerCP/Cy5.5-CD8a, APC/Cy7-CD4, Alexa Fluor 700-CD25, PE-FoxP3, BV605-PD-1, PE-Gr-1, APC-NK1.1, and Pacific Blue Granzyme B were obtained from Biolegend (San Diego, CA). eFluor 450-CD16/32, PE/Cy7-CD206, APC-IFNγ was purchased from eBioscience (San Diego, CA). APC-IL-6 were obtained from thermos fisher (Waltham, MA). BUV737-CD11b, Alexa Flour 700-CD86 were obtained from BD Biosciences (San Jose, CA). PE-Ym1were from Abcam Ltd. (Cambridge, MA). Rabbit anti-Laminin (Cat #L9393) and cariporide (HOE642) was purchased from Sigma Chemicals (St. Louis, MO). Anti-mouse PD-1 (RMP1-14) and IgG2a isotype control were from BioXcell (West Lebanon, NH).

BMT (#B3023), TMZ (#T2577), propidium iodide (PI, #P4864), and MTT (#M2128) were purchased from Sigma-Aldrich (St. Louis, MO). Dulbecco’s Modified Eagle Medium (DMEM/HEPES, Cat# 12430-054) and Penicillin/streptavidin (Cat# 15240062) were from Gibco (Carlsbad, CA). Fetal bovine serum (FBS) was obtained from invitrogen (Carlsbad, CA). Anti-phospho-NKCC1(pThr206) antibody, anti-phospho-SPAK/OSR1 (pSer383 SPAK/pSer325 OSR1) antibody, and anti-total-SPAK/OSR1 (tSPAK/tOSR1) antibody were developed by Dr. Yang (Taiwan National University) and validated in previous studies (Moriguchi et al., 2005 (link); Yang et al., 2010 (link)). Monoclonal anti-total NKCC was from the Developmental Studies Hybridoma Bank (T4, Iowa City, IA). Antibody against α-tubulin (Cat #2125), rabbit anti-phospho AKT (Ser473; Cat# 9271), rabbit anti-AKT (Cat# 4691), rabbit anti-phospho ERK (Thr202/Tyr204; Cat# 4370), rabbit anti-ERK (Cat# 4695), and rabbit anti-phospho p70 S6k (T389; Cat# 9234) were from cell signaling (Beverly, MA). Mouse anti-p70 S6K (Cat# sc-8418) was purchased from Santa Cruz Biotechnology (Dallas, TX). BCA Protein Assay Kit (Cat #23227) was from Thermo Scientific (Rockford, IL). STS66 was synthesized by Töllner et al. (2014) (link) as described previously.