Erk1 2 inhibitor sch772984

CD4⁺ T cells (2 × 10⁴) from HCs with or without anti-CD3/CD28 stimulation were co-cultured with autologous B cells (5 × 10⁴) in 200 μL RPMI 1640 complete medium for 12 days. For SLE samples, freshly purified B cells were co-cultured with autologous CD4⁺ T cells for 12 days. The culture supernatants were collected for IgG detection. The remaining cell mixtures were subjected to phenotypic and functional analysis.
In the blockade assay, function-grade mouse anti-human ICAM-1(eBioscience, San Diego, CA, USA) mAb was added at the concentration of 5 μg/mL in the co-culture system. Erk1/2 inhibitor (SCH772984) (Selleck Chemicals, Houston, TX, USA) was added at the final concentration of 2 μM. PD-L1-Fc protein (Sino Biological, Beijing, China) was added at the final concentrations of 5, 10 and 20 μg/mL in the co-culture.

Antibodies specific for γδ-TCR (GL3), CD27 (LG.3A10), IL-17A (eBio17B7), CD4 (GK1.5), CD8 (53-6.7), CD11b (M1/70), CD11c (N418), IFN-γ (XMG1.2), TNF-α (MP6-XT22), anti-CD3e (2C11) were purchased from BioLegend (San Diego, CA). Recombinant murine IL-1β and IL-23 were from Peprotech (Rocky Hill, NJ). Antibodies specific for p-STAT3, p-STAT5, p-Erk1/2 (Thr202/Tyr204), Erk1/2, p-Jnk1/2 (Thr183/Tyr185), Jnk1/2, and IκB were purchased from Cell Signaling Technology (Boston, MA). The VISTA-specific mAb was as described previously^{8 (link)}. Erk1/2 inhibitor (SCH772984) was obtained from Selleckchem (Houston, TX) and Jnk1/2 inhibitor (SP600125)was obtained from Invivogen (San Diego, CA).