Regulation of IFNα-induced APOBEC3 and IDO by NFκB and calmodulin/CaMK. (A) Whole cell protein lysates collected after treatment with parental IFN or SDM1 (1 ng/mL, 20 minutes) were examined for IκBα phosphorylation by Western blot using an anti–P-IκBα antibody (inset). Transcription of IDO mRNA in macrophages that were pretreated with Bay 11-7082 (10μM) for 1 hour followed by 4 hours of parental or SDM1 mutant is diminished by 82% and 96%, respectively *P < .05 for IDO levels 2C compared with SDM1 and IFN treated compared with inhibitor/IFN treated. Representative data, n = 3. (B) Transcription of APOBEC3A in cells that were pretreated with Bay 11-7082 inhibitor for 1 hour followed by IFNα2c or SDM1 (4 hours), *P < .05 for control compared with IFN-treated; IFNα2c vs SDM1; and IFN constructs treated versus inhibitor/IFN-treated. Representative data, n = 3. (C, D) Macrophage cultures were pretreated for 1 hour with the calmodulin inhibitor W7 (40μM) or (D inset) CaMKII inhibitor KN93 (10-40μM) before 4-hour treatment with parental IFNs and SDM1 mutant. Total mRNA was examined for APOBEC3A and IDO gene transcription, in C P = not significant; in D *P < .05. (E) Whole cell lysates were analyzed for pSTAT1(Y701)(S727) and tubulin (30 minutes). Representative data, n = 3.