Figure 3.

Specific IgG1 class switching inhibition in B cells treated by Iγ1 exon dss ASO. Splenic B cells were isolated from C57BL/6 mice, stimulated with anti-CD40 + IL4 and treated with 2 μM Iγ1 dss ASO (ASO) or an irrelevant ASO (control) for 2 days (B,E) or 4 days (C,D,F,G). (A) Schematic representation of unspliced ε GLT. Iε, ε I exon; Sε, ε switch region; CH1ε, ε constant exon 1; dss, donor splice site. (B,E) Unspliced γ1 (B) and ε (E) GLT expression monitored as described in Figure 2. Iε-for-Q and SεU-rev-Q primers, described in schema A, were used for ε GLT expression determination. (C,F) Post-switch Iμ-Cγ1 and Iμ-Cε mRNA expressions monitored as described in Figure 2. (D,G) Quantification of IgG1 and IgE in culture supernatants by ELISA. (B–G) Data are means ± SEM of two independent experiments, n = 3–4 for each group. Unpaired two-tailed Student's t test was used to determine significance. ns: non-significant, **P <0.01, ***P <0.001.