Figure 3.

Hopx is expressed in naïve B cells. Cells from bone marrow, spleens, and peripheral lymph nodes of Hopx^GFPFoxp3^RFP mice were analyzed by flow cytometry. (A) Representative overlaid histograms show Hopx (GFP) expression among B cells (gated as single, live, CD19⁺, B220⁺) from the bone marrow (blue), spleen (red), or peripheral lymph nodes (gray). (B) Graph shows the percentages of Hopx⁺ cells among B cells from bone marrow, spleens, and peripheral lymph nodes as indicated (n = 4–6 mice from three independent experiments). (C) Representative plot shows Hopx (GFP) expression and anti-IgD staining intensity among splenic B cells. The blue line indicates the cutoff for IgD⁺ and IgD^neg cells further analyzed in D. (D) Graph shows median fluorescence intensity (MFI) of Hopx (GFP) expression among splenic IgD⁺ and IgD^neg B cells as indicated (n = 6 mice from three independent experiments). (E) Representative plot shows Hopx (GFP) expression and anti-CD24 staining intensity among splenic B cells. The blue line indicates the cutoff for CD24^hi and CD24^int cells further analyzed in F. (F) Graph shows median fluorescence intensity (MFI) of Hopx (GFP) expression among splenic CD24^hi and CD24^int B cells as indicated (n = 6 mice from three independent experiments). (B, D, and F) Graphs show mean ± SD. ns – not significant and ∗∗∗∗P < 0.0001 determined by one-way ANOVA with Tukey's multiple comparisons (B) or unpaired two-tailed t test (D and F).