Figure 2: Lsd1 regulates differentiation of TSCs.

(a) Efficiency of Lsd1 knockdown (Lsd1 iKD) and Cre-mediated genetic deletion (Lsd1 iKO) in TSCs is analysed using western blot. Cells were treated with Dox or Tx for the indicated duration. Specificity of Lsd1 knockdown is verified using TSCs expressing short hairpin RNA directed against LacZ (Ctrl iKD). Tubulin is used as a loading control. (b) qRT–PCR analyses of mRNA expression levels of Lsd1 and indicated lineage markers during differentiation in Lsd1 iKO TSCs in the absence or presence of Tx. (c) The expression levels of indicated genes in iKO TSCs transfected with an Lsd1-expression vector (Lsd1), or an empty vector (Ctrl) is monitored during differentiation by qRT–PCR in the absence or presence of Tx. (d) Proliferation assays of Lsd1 iKO TSCs in stemness and differentiation (Diff) conditions in the absence or presence of Tx determined by BrdU incorporation (upper panel) or by cell number using FACS (lower panel). qRT–PCR data are normalized to Gapdh and represented as mean+s.e.m. Experiments (b–d) were independently repeated at least three times in triplicate. Statistical analysis was performed using two-tailed Student’s t-test. *P<0.05; **P<0.01; ***P<0.001.