Figure 4
Rb regulates Elovl6 and Scd1 expression. (a) GSEA result for RB1 target gene set in Rb-depleted MEFs versus control MEFs. (b) GO analysis of upregulated genes in lipid metabolism in Rb-depleted MEFs (upper). Upregulated genes in biosynthesis of unsaturated fatty acids are shown (lower). (c) RT–qPCR of the indicated genes in wild-type MEFs infected with lentiviruses expressing the indicated shRNAs and selected. Cells were cultured in the medium containing 1% FBS for 24 h. Data represent mean+s.d. Four batches of MEFs derived from one embryo were used per one shRNA. *P<0.05, **P<0.01 by Student’s t-test. (d) Immunoblotting (IB) of the indicated proteins in wild-type MEFs transduced with the indicated shRNA and cultured in the medium containing 1% FBS for 24 h. Band intensities were quantified by ImageJ software and indicated. (e) RT–qPCR of the indicated genes in Rb+/+ MEFs and Rb−/− MEFs. Cells were cultured in the medium containing 1% FBS for 24 h. Data represent mean+s.d. Three batches of MEFs derived from each embryo were used per one shRNA. *P<0.05, **P<0.01 by Student’s t-test. (f) Luciferase activity in wild-type MEFs transfected with pGL3 containing the indicated luciferase reporters together with pCMV-β-gal and pCMV vectors expressing the indicated proteins. Results are normalized to β-gal activity. Data represent mean+s.d. (n=3) *P<0.05, **P<0.01 by Student’s t-test. FBS, fetal bovine serum; GO, Gene Ontology; GSEA, Gene Set Enrichment Analysis.
