Fig. 2
Cebpa knockdown up-regulates Ddit3 expression and enhances myeloid progenitor cells susceptibility to ER stress. A Quantitative PCR analysis revealed that the expression of Cebpa was downregulated in 32Dcl3-shCebpa cells compared with the negative control group. B Quantitative PCR analysis revealed that the expression of granulocytic differentiation markers Mpo, Elane and Prtn3 was downregulated in 32Dcl3-shCebpa cells compared with the negative control group. C CFU assays revealed that Cebpa knockdown 32Dcl3 cells developed significantly fewer clones, and rare mature CFU-GM clones could be detected. × 40 magnification. Scale bar, 500 μm. D Quantitative PCR analysis revealed that the expression of Cebpa and granulocytic differentiation markers Mpo, Elane and Prtn3 was continuously inhibited in 32Dcl3-shCebpa cells. E–F 32Dcl3 cells with Cebpa knockdown exhibit elevated mRNA (E) and protein (F) levels of Ddit3 after granulocytic differentiation induced by G-CSF. G 32Dcl3 cells with Cebpa knockdown showed increased apoptotic rate than control cells during granulocytic differentiation induced by G-CSF (100 ng/ml, 48 h). H Quantitative PCR revealed that the mRNA levels of several genes encoding proteins in the pro-survival arm of the UPR were significantly decreased after Cebpa knockdown in 32Dcl3 cells. Data represent Mean ± SD (n = 3); *P < 0.05; **P < 0.01, ***P < 0.001, ****P < 0.0001