Figure 1: Metabolic pathways connected with absent C3ar1/C5ar1 signaling in activated CD4⁺ cells.

A) Sorted CD55^−/−, WT, and C3ar1^−/−C5ar1^−/− CD4⁺ cells were incubated with anti-CD3/28 Dynabeads plus IL-2 for 5 days after which Foxp3⁺ cells were sorted. A) SOCS1, 3, and 4 mRNA levels. Induction in C3ar1^−/−C5ar1^−/− vs WT cells and B) TET1, TET2, and TET3 mRNA levels were quantitated by qPCR. Representative of 2 assays. Induction in C3ar1^−/−C5ar1^−/− vs CD55^−/− cells. C) Sorted Foxp3⁻ CD55^−/−, WT, and C3ar1^−/−C5ar1^−/− CD4⁺ cells were incubated with anti-CD3/28 Dynabeads and IL-2 plus TGF-β in the absence and presence of Vitamin C and percent Foxp3⁺ cells quantitated daily. Induction in C3ar1^−/−C5ar1^−/− vs WT⁻ cells. D) WT CD4⁺ cells were incubated for 1 h with anti-CD3/28 Dynabeads without or with C3ar1/C5ar1 receptor antagonists (RA) or the BRD inhibitor JQ1 after which BRD4, AuraA, AuraB and C3 mRNA levels were assayed by qPCR. Induction in C3ar1^−/−C5ar1^−/− vs WT⁻ cells. All n=2. * = p<.05.