Erdr1 pcmv sport6 plasmid

PerCP-Cy5.5-conjugated anti-CD4 (RM4-5), anti-CD3ε (145-2C11), and anti-hamster IgG1 (G94-56) were purchased from BD Biosciences (San Diego, CA, USA). APC-conjugated anti-CD69 (H1.2F3) was obtained from eBioscience (San Diego, CA, USA). Alexa488-conjugated anti-CD4 (GK1.5) was obtained from BioLend (San Diego, CA, USA). Anti-pPLCγ1 and anti-NFAT1 were obtained from Cell Signaling Technology (Danvers, MA, USA). Anti-PLCγ1 was purchased from Santa Cruz Biotechnology (Dallas, TX, USA).
Recombinant Erdr1 was prepared as described previously [38 (link)]. In brief, the bacterial vector with the Erdr1 CDS region was constructed from the Erdr1-pCMV-SPORT6 plasmid (Open Biosystems, Huntsville, AL, USA), and then Erdr1 was purified with over 95% purity from bacteria. Lots with low endotoxin levels (<0.1 EU/mL) determined by the Limulus Amebocyte lysate assay (Cape Cod, East Falmouth, MA, USA) were used for experiments.

Alexa488-conjugated anti-Foxp3 (MF23), PerCP-Cy5.5-conjugated anti-CD4 (RM4-5), FITC-conjugated anti-CD25 (3C7), PE-conjugated anti-CD138 (281-2), and anti-CD3ε (145-2C11) were purchased from BD Biosciences (San Diego, CA, USA). PE-conjugated anti-CTLA4 (UC10-4B9), APC-conjugated anti-Foxp3 (FJK-16s), and APC-conjugated anti-CD69 (H1.2F3) were obtained from eBioscience (San Diego, CA, USA).
Recombinant Erdr1 was prepared as described previously [32 (link)]. Briefly, the bacterial vector with the CDS region of Erdr1 was prepared from the Erdr1-pCMV-SPORT6 plasmid (Open Biosystems, Huntsville, AL, USA). Erdr1 with more than 95% purity was purified from bacteria. The endotoxin level was examined by the Limulus amebocyte lysate assay (Cape Cod, East Falmouth, MA, USA). Low endotoxin (<0.1 EU/mL)-contained lots were used for experiments.