Fig. 5
From: mTOR intersects antibody-inducing signals from TACI in marginal zone B cells
TACI triggers CSR by cooperating with TLR9 through mTORC1. a PCR analysis of Iγ1/2-Cγ1, Iγ1/2-Cγ2, Iγ3-Cγ3, Iα1-Cα1, Iα2-Cα2 and Iµ-Cµ germline transcripts (GTs) as well as Iγ1/2-Cµ, Iγ3-Cµ, Iα1/2-Cµ switch circle transcripts (SCTs) RT-PCR-amplified from human splenic MZ B cells incubated with APRIL in the presence or absence of rapamycin (rapa) for 3 days. Red numbers indicate band intensity relative to Iµ-Cµ bands. b qRT-PCR of mRNA for AID (AICDA) from human splenic MZ B cells incubated with APRIL in the presence of control vehicle (vehi) or rapamycin for 3 days. Results are normalized to β-actin mRNA and presented as relative expression (RE) compared to B cells incubated with medium alone (Ctrl). c IB of AID and β-actin from human splenic MZ B cells treated as in a. Red numbers indicate band intensity relative to β-actin. d GSEA showing enrichment or depletion of coordinated proliferation-related gene sets from human splenic MZ B cells cultured for 3 h with or without APRIL or rapamycin. Ctrl, medium alone. NES indicates correlation between individual gene sets. Positive correlation, NES > 0; negative correlation, NES < 0. Heat map showing top 25 genes similarly enriched or depleted in MZ B cells exposed to different treatments. Colors correspond to significant FC expression: red, high expression; blue, low expression. Bold indicates gene products discussed in the text. Data represent one of three experiments with similar results (a, c), summarize three experiments (b) or correspond to one experiment with three biological replicates (d). Error bars, s.e.m.; *p < 0.05, **p < 0.01 (two-tailed Student’s t test)
