Fig. 3
From: GDF11 expressed in the adult brain negatively regulates hippocampal neurogenesis
Construction and characterization of Gdf11cKO mice. A Schematic of Gdf11 flox (top), and Gdf11 ∆ (bottom) alleles. Primer binding sites are labeled by a, b, and c alongside their amplicon sizes. B Genomic DNA PCR for Gdf11 flox (b-c amplicon: 389 bp) and Gdf11 ∆ (a-c amplicon: 330 bp) amplicons from representative Gdf11flox/flox and Gdf11cKO mice. These data confirm recombination of the Gdf11 floxed allele in the Gdf11cKO, but not Gdf11flox/flox, mice. C Quantitative PCR analysis of Gdf11 expression in whole brain, hippocampus, cerebellum, kidney, heart, spleen, lung, muscle (tibialis anterior), and liver tissue isolated from Gdf11flox/flox and Gdf11cKO mice. Hprt was used as a housekeeping gene. Relative mRNA levels for all samples are normalized to Gdf11flox/flox mice and error bars depict SEM. D Concentration of GDF11 protein in serum of Gdf11flox/flox and Gdf11cKO mice at 3 months old, as detected by mass spectrometry (n = 9 mice per genotype, males and females pooled.) Individual data points are overlaid with mean ± SEM. E RNAscope in situ hybridization micrographs of mouse hippocampi (dentate gyrus) probed for Gdf11 (white puncta) from representative Gdf11flox/flox and Gdf11cKO mice. Scale bar = 20 μm. F Scatterplot showing the quantification of RNAscope data (data includes mean of 4 control and 4 Gdf11cKO brains). G Quantitative PCR analysis in isolated hippocampi from Gdf11flox/flox and Gdf11cKO mice. Analysis of TGFβ family members Mstn, Tgfβ1, Tgfβ2, Tgfβ3 expression and GDF11 receptors Alk4 and Alk5 expression. Relative mRNA levels for all samples were normalized to the Gdf11flox/flox group and error bars depict SEM. No statistically significant difference between groups was observed for any GDF11 family member or receptor studied. H Body weight (grams) measured for male and female Gdf11flox/flox or Gdf11cKO mice over varying lengths of time post 3 consecutive days of single tamoxifen IP injections (0.2 mg/gram b.w.). All statistics were calculated using two-tailed t-test. *p < 0.05, **p < 0.01