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. 2016 Dec 12;213(13):3007–3024. doi: 10.1084/jem.20160712

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© 2016 Hayakawa et al.

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Figure 7. — Continued up-regulated c-Myc and down-regulated Bmf by B1 B cells to become CLL. (A) CXCR5 up-regulated B1a cell migration, interaction, and renewal/maintenance from young to aged, with cyclin D2 dependence. (B) Myc mRNA levels by B1 B cells at normal and tumor stage, including TC^– ATA B cell tumors, relative to FO B cells. Mean percentages indicated in lines. S, spleen; p, peritoneal cavity. (C) Reduction of Myc mRNA by normal mouse (C.B17) B1a during 24-h culture. After plating in a 96-well U-plate for culture (with 10% FCS), the immediately harvested samples before 37°C incubation was defined as 0 h. FO B Myc mRNA levels were similar before and after culture (as 1.0). n = 4; mean ± SE; *, P = 0.0007. (D) Myc, Mcl-1, Bcl-2, Bim, and Bmf mRNA levels by B1 B cells at TC^– nontumor and TC⁺ (and TC^–) tumor stage relative to 2-mo-old FO B cells. (left) WT (C.B17 mice). (right) ATA B cells in ATAμκTg mice, including arrested ATA B cells in spleen. Values are mean ± SE. (E) Western blot of samples of FO B and pB1a cells in 2-mo-old WT mice together with TC⁺ CLL/lymphomas stained for Myc, Mcl-1, Bcl-2, Bim, Bmf, and β-actin. Another set of WT/ATA B cell tumors was assembled to compare Myc, Bim, Bmf, and β-actin. WT FO B and pB1a sample data were similar to WT data on left (not depicted). Representative data of two to three experiments. (F) Western blot using rabbit anti-Bmf, in comparison with Myc. (G) Myc, Bim, and Bmf mRNA levels in immature B cells (CD19⁺B220⁺AA4⁺IgM⁺IgD^–) in day 1 liver and 2 mo BM. 2-mo-old spleen FO B cells in 1.0. C.B17 mouse. n = 4; mean ± SE; *, P = 0.016; **, P = 0.001.