Fig 1: Effects of MIA-690 (5 µg) and MR-409 (1-5 µM) on IGF-1 levels in serum of DSS-treated mice (n = 6 for each group of treatment) as determined by ELISA kit. Data are expressed as means ± S.E.M. and analyzed by analysis of variance (ANOVA) followed by Bonferroni post-hoc test *p < 0.05 versus water controls, DSS or DSS + MR-409 treated mice.
Fig 2: Schematic presentation of a working model for the role of PON2 in ovarian tumor growth.ID8 cells form tumors in immunocompetent C57BL6/J mice. ID8hPON2 cells overexpressing human PON2 develop reduced tumors. Based on the data presented in this paper, PON2 overexpression regulates three anti-tumorigenic pathways. I). PON2 overexpression reduces mitochondrial superoxide levels, which regulate c-Jun activation. Reduced c-JUN binding to the IGF-1 promoter leads to decreased expression of IGF-1 protein. II). PON2 expression enhances electron transport chain activity leading to decreased cholesterol levels resulting in impaired caveolin-1 and IGF-1R interaction. I & II result in decreased cell proliferation and reduced tumor growth
Fig 3: Overexpression of hPON2 inhibits cell proliferation via IGF-1 signaling pathway by altering the caveolin-1 and IGF-1R interaction in ID8 cells.a) 5 × 103 cells were plated in 96 well plates and were incubated at 37 °C in 5% CO2. After 24 h, cells were serum starved for 12 h. Following this, cells were treated with IGF-1 at the concentration of 500 nM or vehicle for 6 h, 12 h, and 24 h. Cell proliferation was measured as described above. b) ID8EV cells or ID8hPON2 cells were cultured in six-well plates as described above and treated with IGF-1 at the concentration of 1 µM for 30, 60, and 90 min. 50 µg of total protein (100 µg for ERK1/2) was subjected to western blotting and phosphorylated IGF-1R, total IGF-1R, phosphorylated ERK1/2, total ERK1/2, cyclin D1, and vinculin were detected using respective antibodies as described in the materials and methods section. c) Immunoprecipitation was performed with caveolin-1 antibody and IGF-1R was detected by western blotting. d) Lipids were extracted from ID8EV and ID8hPON2 cells using chloroform: isopropanol: NP (7:11:0.11), air dried at 50 °C to remove the chloroform and placed under vacuum for 30 min, dispersed with assay buffer and cholesterol was measured as described in the Materials and methods section. e) Mitochondria and cytosol were isolated as described in the materials and methods section and acetyl CoA was measured using a kit according to manufacturer’s protocol. Values were normalized based on protein concentration. Values were expressed as fold change (n = 3). f) Citrate lyase enzyme activity was measured as described in the Materials and methods section. (n = 3). *p < 0.05, compared to ID8EV
Fig 4: Acu-LFES upregulates IGF-1 mRNA and protein in the muscle of diabetic mice.Panel A: Total RNA isolated from combined gastrocnemius and EDL muscles of control, Acu-LFES, diabetes or diabetes/Acu-LFES mice were assayed for IGF-1 expression by real time qPCR. The bar graph shows mRNA from the muscles of each group of mice. Results are normalized to 18S RNA (Bars: mean ± s.e.; n = 9/group; * = p<0.05 vs. control and # = p<0.05 vs. diabetes). Panel B: IGF-1 protein levels were measured by ELISA in in combined gastrocnemius and EDL lysates from control, Acu-LFES treated normal controls (Acu-LFES), diabetes or diabetes/Acu-LFES mice. The bar graph shows the IGF-1 protein levels in each group. IGF-1 levels in the muscle lysates were normalized to the total protein concentration (Bars: mean ± s.e.; n = 9; * = p<0.05 vs. control and # = p<0.05 vs. diabetes).
Fig 5: Male offspring from prazosin treated dams display characteristics of dwarfism. Body length was measured at the age of 4–5 months, and male prazosin offspring were significantly shorter when compared to age matched control animals (A). IGF-1 serum levels were decreased (B; control: N = 4, prazosin: N = 8), total T4 levels normal (C), and gene expression analysis of pituitary (D) and liver (E) mRNA revealed changes in growth hormone receptor (Ghr) expression. Hepatic DNA methylation of selected CpG sites in the Igf1 (F) and Ghr (G) gene as assessed by bisulfite pyrosequencing (H). Hepatic Ghr mRNA expression and CpG -13488 DNA methylation for all animals was inversely correlated (I). N = 4–8 animals per group (B) and N = 8 animals per group (A, C–I). All values are mean ± S.E.M. and P < 0.05 (*), P < 0.01 (**) or P < 0.001 (***) with Student's t-test and Welch's correction (A–E) or Holm-Sidak correction for multiple testing (H) or Pearson's correlation (I). ATG: translational start; AU: arbitrary unit; Dio1: deiodinase 1; Gh: growth hormone; Ghr: growth hormone receptor; Igf1: insulin-like growth factor 1; Igfbp3: insulin-like growth factor binding protein 3; PPARg: peroxisome proliferator-activated receptor gamma; Prl: prolactin; Tshb: thyroid stimulating hormone b; TSS: transcriptional start site.
Supplier Page from Abcam for Mouse IGF1 ELISA Kit