Figure 7
From: Baf53a is involved in survival of mouse ES cells, which can be compensated by Baf53b
Baf53a- or Baf53b-expressing Baf53a-deficient ES cells exhibit the undifferentiated state. (A) Alkaline phosphatase staining of Baf53a- or Baf53b-expressing Baf53a-deficient ES cells. Baf53a cKO ES cells were transfected with expression vectors for empty (ev), Baf53a, or Baf53b. Transfected ES cells were divided into two dishes at 48 h after transfection and cultured with or without 1 μg/mL Tet. Alkaline phosphatase activity of these ES cells was detected by the VECTOR Red alkaline phosphatase substrate kit at 6 days after Tet treatment. Scale bar is 100 μm. (B) Expression of markers of undifferentiated cells in Baf53a- or Baf53b-expressing Baf53a cKO ES cells. Expression of Oct3/4, Nanog, Esrrb, Klf4, Rex1, and Dax1 mRNA in the indicated samples was examined by quantitative RT-PCR after 6 days. Because the Tet-treated, empty vector-transfected Baf53a cKO ES cells died after 6 days, mRNA expression was not examined. (C) Expression of p21 mRNA in Baf53a- or Baf53b-expressing Baf53a cKO ES cells. Expression of p21 mRNA in the indicated samples was examined by quantitative RT-PCR. The data in panels B and C were normalized to GAPDH expression. *p < 0.05 (t-test). (D) Growth curves of Baf53a- or Baf53b-transfected Baf53a-deficient ES cells. Baf53a- or Baf53b-transfected Baf53a-deficient ES cells were continually cultured in the presence of Tet and LIF and cell numbers were counted over time. The representative results shown are expressed as the means ± standard deviation of three independent experiments. Note: OE = overexpression. (E) DNA microarray analysis of Baf53a- and Baf53b-expressing, Baf53a-deficient ES cells. Scatter plots of log-ratios of relative expression levels were shown.
