Comparative RPA analysis of cytokine mRNA expression of BM DCs terminally differentiated by LPS or CD40 ligation.

(A) Comparison of cytokine mRNA expression in FACS-sorted CD86⁺ BM DCs (control) and DCs terminally differentiated with LPS or anti-CD40 IgM mAb. Expression of IFN-α mRNA was also investigated in CD86⁺ BM DCs infected with a rAd encoding no transgene (MOI = 100) used as positive control. mRNA was analyzed by RPA. DCs treated with irrelevant IgM exhibited a pattern similar to that of the control group (not shown). Data are from a single experiment representative of 3 separate experiments. (B) Quantitative analysis of mRNA cytokine gene expression. Densitometric analysis of each lane was performed on scanned autoradiographs, and all values are expressed relative to corresponding housekeeping gene transcripts (L32). ▪, control DC; ■, DC + LPS; ░, DC + anti-CD40. Densitometric values (means ± 1 SD) were pooled from 3 separate experiments.