Fig. 4: The regulatory activity of miR-155 is revealed under hypoxia.
From: Epi-microRNA mediated metabolic reprogramming counteracts hypoxia to preserve affinity maturation
a Representative flow cytometric histograms of hypoxia dye, MAR vs. relative cell number (RCN) (left). Relative MFI of MAR (right) in GC-B and non-GC-B cells. GC-B and non-GC-B cells were isolated from inguinal lymph nodes of C57BL/6 mice. One-way ANOVA. Pooled from three independent experiments (n = 6). b KDM2A MFI of B cells from WT and KO mice cultured under normoxia (21% O2) or hypoxia (1% O2) for the indicated time periods. Unpaired student’s t-test, two-tailed. Pooled from three independent experiments (WT n = 5; KO n = 5). c Representative flow cytometric histograms of CTV vs. RCN at day 2 and 4 (top). CTV MFI of B cells from WT and KO mice cultured under normoxia or hypoxia for the indicated time periods (bottom). Unpaired student’s t-test, two-tailed. Pooled from five independent experiments (WT n = 5; KO n = 5). d Representative image of B cells from WT and KO mice cultured under normoxia or hypoxia for two days, followed by staining with MitoTracker Deep Red (Red) and MitoTracker Orange CM-H2 TMRos (green). DAPI (blue) was used as a nuclear counterstaining. Scale bar, 10 µm. Representative images are shown. e Quantification of relative MitoTracker Orange CMTMRos levels. Each dot represents one cell. One-way ANOVA. Pooled from two independent experiments (WT n = 4; KO n = 4). f Relative ATP levels in B cells from WT and KO mice. The replated cells were cultured for two hours in hypoxia with indicated media for ATP measurement. The luminescence values of each type of B cells cultured in media containing glucose for two hours in hypoxia were used to calculate relative ATP levels. One-way ANOVA. Pooled data from three experiments (WT n = 3; KO n = 3). Unless otherwise stated, mean ± SEM is indicated. n.s., not significant.
