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Blood, Vol. 111, Issue 3, 1396-1403, February 1, 2008
Interferon regulatory factors 4 and 8 induce the expression of Ikaros and Aiolos to down-regulate pre–B-cell receptor and promote cell-cycle withdrawal in pre–B-cell development
Blood Ma et al.
111: 1396
Supplemental materials for: Ma et al
Files in this Data Supplement:
- Table S1. Sequences of the primer sets (JPG, 75.5 KB)
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- Figure S1. IRF4ER or IRF8ER can rescue the development of IRF4,8−/− pre-B cells (JPG, 70.8 KB)
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IRF4,8−/− pre-B cells were infected with virus expressing IRF4ER and IRF8ER. 24h after infection, cells were treated with either placebo (ethanol only) or Tamoxifen (1µM) and cultivated in the absence of IL7 for another 36h. The expression of Kappa and CD25 was examined by FACS in the presence or absence of Tamoxifen. The numbers represent the percentages of the gated cells.
- Figure S2. Ikaros DN blocks Aiolos induced cell cycle withdrawal in IRF4,8−/− pre-B cells (JPG, 55.4 KB)
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IRF4,8−/− pre-B cells were co-infected with Aiolos (YFP) and IkarosDN (GFP). The infected cells were stained with Hoechst dye and the cell cycle status of the infected cells was analyzed three days later using strategies described in Fig. 5.
- Figure S3 (JPG, 53.3 KB)
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(A) IRF4,8−/− pre-B cells are hypersensitive to IL7 stimulation and reconstitute expression of IRF4, Ikaros or Aiolos does not affect the expression of IL-7 receptor alpha chain (IL-7R). B220+ cells were isolated from the bone marrow of IRF4+/−IRF8+/− and IRF4,8−/− mutant mice. The cells were cultivated on top of an irradiated S17 stromal layer in the presence of IL-7. The cells at 105 / well were plated into a 96-well plate in the presence of different concentrations of IL-7 as indicated. The cells were pulse-labeled with 3H-Thymidine for 6 hours and the radioactivity was measured by a Microplate scintillation counter. (B/C) The RNAs isolated from the infected cells described in Fig. 1C and Fig. 3B were used for real-time PCR analysis to detect the expression of IL-7R.
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