Figure 6.
Figure 6. Reduced erythropoietic supporting ability of Ezh2-inactivated adult BM stromal cells. (A) Mmp9 mRNA expression level in DMSO and 10 μM GSK126 (Ezh2 inhibitor) treated MS-5 cells (1 experiment). (B) ChIP-qPCR for Ezh2, H3K27me3, and H3K27ac at Mmp9 locus in DMSO or GSK126 treated MS-5 cells (3 independent experiments). (C) In vitro coculture assay of MS-5 cells treated with DMSO, GSK126, and/or Mmp9 inhibitor (2 independent experiments). Numbers of replicates are indicated at the bottom of each column. Two-tailed Student t tests were used to assess statistical significance. Error bars represent SEM. (D) Representative immunofluorescence staining (n ≥ 30 cells, 2 independent experiments) of MS-5 cells collected post in vitro treatment with DMSO, GSK126, and/or 10 nM Mmp9 inhibitor using antibodies against mKitL (green). Nuclei were stained with DAPI (blue). Scale bars, 20 µm.

Reduced erythropoietic supporting ability of Ezh2-inactivated adult BM stromal cells. (A) Mmp9 mRNA expression level in DMSO and 10 μM GSK126 (Ezh2 inhibitor) treated MS-5 cells (1 experiment). (B) ChIP-qPCR for Ezh2, H3K27me3, and H3K27ac at Mmp9 locus in DMSO or GSK126 treated MS-5 cells (3 independent experiments). (C) In vitro coculture assay of MS-5 cells treated with DMSO, GSK126, and/or Mmp9 inhibitor (2 independent experiments). Numbers of replicates are indicated at the bottom of each column. Two-tailed Student t tests were used to assess statistical significance. Error bars represent SEM. (D) Representative immunofluorescence staining (n ≥ 30 cells, 2 independent experiments) of MS-5 cells collected post in vitro treatment with DMSO, GSK126, and/or 10 nM Mmp9 inhibitor using antibodies against mKitL (green). Nuclei were stained with DAPI (blue). Scale bars, 20 µm.

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