Supplementary Figure 1: Quantitative real-time PCR analysis of membrane IgG1 and membrane IgE transcripts in IgG1+ GFP− (IgG1), IgG1+ GFPhi (DP), and IgG1− GFPhi (IgE) cells sorted from pooled mesenteric and mediastinal lymph nodes of IgE-GFP reporter mice 15 days after infection with N. brasiliensis.
From: Addendum: IgE+ memory B cells and plasma cells generated through a germinal-center pathway
(a) Membrane IgG1 transcript. (b) Membrane IgE transcript. Cells were first gated as B220+ IgD− B cells, then as GL7+ CD95+ CD38− germinal center B cells, and then separated based on IgG1 and GFP expression. The location and sequences of primers and probes for the membrane IgG1 and membrane IgE assays are as shown in the figure. Approximate cell input for each assay was as follows: 4×104 IgG1 cells, 1×104 DP cells, and 1×104 IgE cells. Data are plotted as relative expression, with the normalized cycling threshold (CT) value denoted above each bar graph. For each assay, results were normalized to those of control transcription of the gene encoding ribosomal protein L19, and relative transcription was determined by the calculation 2−ΔCT with reference to an absolute baseline CT of 40 cycles. The graph depicts the mean ± s.e.m. of two technical replicates.
