Restorative effect of Angptl3 on functional deficiency of HSCs from the ATF4^−/− FL. (A) Representative flow cytometric analysis of LSK cells cultured in the presence or absence of Angptl3 for 4 days (3 independent experiments). (B) Fold change of cell number and percentage of LSK and CD48^–LSK cells after culturing ATF4^−/− HSCs with or without Angptl3 for 4 days (3 independent experiments). (C) Donor chimerism of the ATF4^−/− SLAM cells cultured with or without Angptl3 for 4 days at 16 weeks after transplantation (3 independent experiments). (D) Percentage of input of the ATF4 antibody using mouse FL cells based on the chromatin immunoprecipitation (ChIP) assay. Primers were designed to amplify the Angptl3 promoter (n = 3 for each group). Expression of (E) Angptl3 mRNA or (F) protein by transfecting ATF4 siRNA into an NIH/3T3 cell line (3 independent experiments). (G) Expression of Angptl3 mRNA by transfecting ATF4 complementary DNA (cDNA) overexpression vector into an NIH/3T3 cell line (4 independent experiments). Student t test *P < .05, **P < .01, and ***P < .001. Ab, antibody; IgG, immunoglobulin G; MIG, mouse stem cell virus-internal ribosome entry site-green fluorescent protein.