ArrayExpress Uploader07419PGC-1alpha -/-control siRNAPGC-1alpha -/-control siRNAPGC-1alpha -/-PGC-1beta siRNA knockdownPGC-1alpha -/-PGC-1beta siRNA knockdownwild_typeuntreatedwild_typeuntreatedarrayexpress_sid6Mitochondria play an essential role in the ability of brown fat to generate heat, and the PGC-1 coactivators control several aspects of...16679291E-GEOD-5042/profile/8773/arrayexpressuploader26adipose tissuebrown adipose tissueDec.12, 2014Falsetranscription-profiling-of-differentiated-brown-adE-GEOD-5042_A-AFFY-45Transcription profiling of differentiated brown adipocytes from wild type, PGC-1 alpha knockout, or PGC-1 alpha knockout and PGC-1 beta knockdown mice to investigate the roles of PGC-1 coactivators in brown fat differentiation.Nov.12, 2014Mitochondria play an essential role in the ability of brown fat to generate heat, and the PGC-1 coactivators control several aspects of mitochondrial biogenesis. To investigate their specific roles in brown fat cells, we generated immortal preadipocyte lines from the brown adipose tissue of mice lacking PGC-1±. We could then efficiently knockdown PGC-1β expression by shRNA expression. Loss of PGC-1± did not alter brown fat differentiation but severely reduced the induction of thermogenic genes. Cells deficient in either PGC-1α or PGC-1β coactivators showed a small decrease in the differentiation-dependant program of mitochondrial biogenesis and respiration; however, this increase in mitochondrial number and function was totally abolished during brown fat differentiation when both PGC-1± and PGC-1 were deficient. These data show that PGC-1± is essential for brown fat thermogenesis but not brown fat differentiation, and the PGC-1 coactivators play an absolutely essential but complementary function in differentiation-induced mitochondrial biogenesis. Affymetrix microarray analysis of total RNA from wt, PGC-1± KO and PGC-1± KO; cells expressing an RNAi specific for PGC-1 knockdown was performed. Of the 461; mitochondrial genes analyzed, 181 were found to be at least 20% different between wt; and defective PGC-1± and β adipocytes (p < 0.05). More than 85% of these genes were downregulated in cells deficient for PGC-1alpha and PGC-1beta. Experiment Overall Design: Brown preadipocytes that were either WT, KO for PGC-1alpha, or KO for PGC-1alpha and deficient for PGC-1beta (knockdown through siRNA expression) were differentiated for seven days. RNA was made from biological replicates of the three different types of brown adipocytes (WT, KO expressing a control siRNA, KO expressing a siRNA specific for PGC-1beta knockdown).http://www.ebi.ac.uk/arrayexpress/experiments/E-GEOD-5042mousehttp://www.ebi.ac.uk/arrayexpress/experiments/E-GEOD-5042/samples/