El4 atcc tib 39 cells

EL4 (ATCC® TIB-39™) cells were purchased from ATCC. The hybridoma RMP1-14 for αPD-1 production was provided by Professor Hideo Yagita at the Juntendo University School of Medicine. DH5α competent E.coli cells were purchased from Thermo Fisher Scientific Inc. (Waltham, MA). SHuffle® T7 Competent E. coli cells were purchased from New England Biolabs (Ipswich, MA). Expression vector pET-25b(+) was purchased from EMD Millipore (Billerica, MA). Restriction endonucleases were purchased from New England Biolabs (Ipswich, MA). LB media were prepared in our lab using the standard formula. Cell culture media and supplements including RPMI-1640, Dulbecco's Modified Eagle Medium (DMEM), and fetal bovine serum (FBS) were purchased from Thermo Fisher Scientific Inc. (Waltham, MA). B6.129S7-Rag1^tm1Mom/J mice and NOD/ShiLtJ mice were purchased from the Jackson Laboratory. All the animal experiment protocols were approved by the Institutional Animal Care and Use Committee at the University of Utah.

Female C57BL/6 mice were purchased from The Jackson Laboratories. Animal studies were conducted following a protocol approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Utah. EL4 (ATCC^® TIB-39^™) cells were purchased from ATCC and were maintained in DMEM medium with 10% horse serum. EL4 (PD-1^KO) cells were generated according to our previous research and maintained in the same medium for EL4 [15 (link)]. Macrophage Raw 264.7 cells were purchased from ATCC and maintained in RPMI 1640 medium with 10% FBS. Expi293 expression system was purchased from ThermoFisher and applied following the manufacture instruction. PE anti-mouse IgG2a antibody (clone: m2a-15F8) was purchased from Ebioscience. APC anti-mouse CD3 (clone: 17A2), FITC anti-mouse CD3 (clone:17A2), BV510 anti-mouse CD4 (clone: RM4–5), APC/Cyanine7 anti-mouse CD8 (clone: 53–6.7), PE anti-mouse TCR Vβ12 (clone: MR11–1), APC anti-mouse PD-1 (homemade), APC anti-mouse CD11b (clone: M1/70), PE anti-mouse PD-1 (clone: RMP1–14) and PE anti-mouse FcγRIV (clone: 9E9) were purchased from Biolegend.