Fig 1: Genome-wide screening and genetic basis of PDGFD for tail configuration.a Different phenotypes in tail configurations; picture credit: Xin Li. b, c Statistic VST is plotted for selected CNVs through pairwise comparison on chromosomes 7 b and 14 c with same threshold VST value = 0.64. d Selective regions associated with tail configuration by XP-CLR using the SNP data with the threshold XP-CLR = 8.26. Candidate genes overlapping with the regions, which are significantly selected by XP-CLR & ln(p ratio)/ln(2), XP-CLR & ln(p ratio)/ln(2) & iHS, and XP-CLR & ln(p ratio)/ln(2) & HKA are marked by gray, orange, and blue colors, respectively. Below this plot, genes near the peaks are indicated by green boxes. The pie charts represent the spectrum of allele frequencies at the non-synonymous loci of PDGFD in populations of different tail configurations. The type of variant allele is indicated in blue, while the reference allele in pink. e Genotype patterns for the promoter region of PDGFD among 11 fat-tailed/rumped, 11 thin-tailed sheep, and Asiatic mouflon. f Structures and expression levels of four isoforms of PDGFD. Expression levels are shown in varying shades of yellow color. g, i Expression pattern of control gene ß-actin and target gene PDGFD in tail fat examined by RT-PCR g and western blot analysis i. h, j The relative expressions of PDGFD in tail fat by real-time PCR (qPCR) h and western blot analysis j. k Adipogenesis signaling pathway46 and the inhibitory function of PDGFD in differentiation of white adipocytes45 by activating PDGFRß signaling44. All experiments were repeated three times with similar results. Samples derived from the same experiment and the blots were processed in parallel. g–j Experiments were performed with the control sample (the thin-tailed sheep; MFW) and target samples (long fat-tailed sheep (HDW), fat-rumped sheep (ALS) and short fat-tailed sheep (SXW)). The data in h and j are presented as the mean ± SD, n = 3 biologically independent samples; groups with significant differences (*P < 0.05; **P < 0.01) were performed by two-tailed unpaired t-test. Source data are provided as a Source Data file.
Fig 2: IL-15 induces PDGF-D expression in an autocrine manner. (A) mRNA levels of PDGFA, PDGFB, PDGFC, and PDGFD in T cells, B cells, and NK cells were analyzed using the BioGPS online tool. (B) mRNA levels of PDGFA, PDGFB, PDGFC, and PDGFD in T cells, B cells, and NK cells were examined by qPCR (n = 10). (C) Primary NK cells were treated with IL-15 (10 ng/mL) for the indicated times. mRNA levels of PDGFD were examined by qPCR (n = 3). (D and E) Representative dot plots and percentages of PDGF-D levels in NK cells after IL-15 (50 ng/mL) treatment for 24 h. Resting NK cells were used as control (n = 5). (F) Immunoblotting shows the full length and cleavage of PDGF-D in resting and IL-15-treated NK cells. (G) ELISA shows PDGF-D levels in supernatants of NK cell cultures (n = 6). (H) Luciferase reporter assay shows that p65 activates PDGFD gene transcription. (I and J) Binding of p65 to the PDGFD promoter in IL-15–treated (I) or resting NK cells (J) as determined by ChIP-qPCR (n = 3). Data represent three independent experiments. Data shown are means ± SD. NS, not significant, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.
Supplier Page from Abcam for Anti-SCDGFB/PDGF-D antibody [EPR7266(2)]