Anti PPAR γ 1 + 2  

Western Blot of Rabbit Anti-PPAR gamma 1&2 Antibody. Lane 1: Opal Prestained Molecular Weight Marker (p/n MB-210-0500). Lane 2: rec PPAR protein [0.05μg]. Lane 3: rec PPAR protein/HeLa Whole Cell Lysate [0.05μg/10μg]. Primary Antibody: Anti-PPAR gamma 1&2 at 1.0μg/mL overnight at 2-8°C. Secondary Antibody: Goat Anti-Rabbit IgG HRP (p/n 611-103-122) at 1:70,000 for 30mins at RT. Block: BlockOut Buffer (p/n MB-073). Predicted MW: rec. protein (54.5kDa), Isoform 1 (54.6kDa), Isoform 2 (57.6kDa). Observed MW: ~63kDa.

Impaired protein expression of miR-143 target genes with aging in preadipocytes before and after adipogenic differentiation. A decreased adipogenic differentiation potential of preadipocytes from old rats (30 mo) compared to 6 mo (n？=？8/group) rats was due to changes in miR-143 levels. Western blot was used to analyze the protein levels of miR-143 target genes, ERK5,？5A？and？B; PPARg,？5A？and？C; or osteogenic marker, Runx2,？5A and D？in the young and old rats preadipocytes before and after adipogenic differentiation. RT-qPCR was also used to measure adipogenic markers, adiponectin,？5E？and IL-6,？5F. The data was expressed as ratio or fold of relative expression ±SEM (Standard Error of Mean). *: p<0.05; **: p<0.01. Fig 5. PMID:？23516615

？Gain and loss of function assays to validate the role of miR-143 in young (6 mo) and old (30 mo) rat SVF cells, pre-adipogenesis. The gain or knockdown of miR-143 by premir or antagomir-143 was performed by transient transfection of both young and old rat (n？=？8) preadipocytes, pre-adipogenesis. The transfection efficiency and target gene levels were confirmed by RT-qPCR and Western blot analysis. miR-143,？7A; ERK5 mRNA,？7B; PPARg mRNA,？7C; IL-6 mRNA, 7D; ERK5 protein,？7E; and PPARg protein,？7F.？The real time PCR data was expressed as fold of relative expression ±SEM (Standard Error of Mean). *: p<0.05; **: p<0.01. Fig 7. PMID:？23516615