Journal: Andrology

Article Title: Catch-up fat in male adults induces low testosterone and consequently promotes metabolic abnormalities and cognitive impairment.

doi: 10.1111/andr.13177

Figure Lengend Snippet: FIGURE 3 Alteration of cognition function, insulin signal pathway, and NR2A/2B expression in the cerebral cortex of male catch-up fat in adult (CUFA) rats. Comparison of the numbers of trials required to reach the criterion of six consecutive correct in the different stages of ASST (A), the relative mRNA expression level of IRS-1 (B), insulin-stimulated AKT phosphorylation at Ser473 (C), the relative protein expression level of AR (D), relative mRNA expression (E) and protein expression (F) level of NR2A, relative mRNA expression (G) and protein expression (H) level of NR2B between NC and RN groups at 12 w. *P < 0.05, **P < 0.01, ***P < 0.005. Abbreviations: NC, normal chow controls; RN, caloric restriction and refeeding with normal chow (CUFA rats); ASST, Attentional set-shifting task; SD, Simple discrimination; CD, Compound discrimination; R1, First reversal; IDS, Intra-dimensional set-shifting; R2, Second reversal; EDS, Extradimensional set-shifting; R3, Third reversal; IRS-1, Insulin receptor substrate 1, AKT, serine/threonine kinase 1; AR, androgen receptor; NR2A, N-methyl-d-aspartate receptors subtype 2A, NR2B, N-methyl-d-aspartate receptors subtype 2B

Article Snippet: Proteins extracts from the cerebral cortex of rats were prepared and immunoprecipitation was performed as reported previously.3 The antibodies used for immunoblot assays were anti-PKB (Protein kinase B)/Akt (1: 1000, Cell Signaling Technology), anti-phosphoPKB/Akt (Ser473) (1: 1000, Cell Signaling Technology), anti-NR2A (1:2000, Proteintech Group, Inc.), anti-NR2B (1:2000, Proteintech Group, Inc.), anti-AR (androgen receptor) (Proteintech Group, Inc), and anti-GAPDH (1:3000, Gene Tex).

Techniques: Expressing, Comparison, Phospho-proteomics

Journal: Andrology

Article Title: Catch-up fat in male adults induces low testosterone and consequently promotes metabolic abnormalities and cognitive impairment.

doi: 10.1111/andr.13177

Figure Lengend Snippet: FIGURE 6 Testosterone supplementation attenuated cognitive impairment as well as the impaired insulin signal pathway and suppressed NA2A/2B expression in male catch-up fat in adults (CUFA) rats. Comparison of the number of trials required to achieve the criterion of six consecutive correct in the different stages of ASST (A), the relative mRNA expression level of IRS-1 (B), insulin-stimulated AKT phosphorylation at Ser473 in brain tissue (C), the relative protein expression level of AR (D), relative mRNA expression (E) and protein expression (F) level of NR2A, relative mRNA expression (G) and protein expression (H) level of NR2B between RN and RT groups at 12w. *p < 0.05, **p < 0.01, ***p < 0.005. Abbreviations: RN, caloric restriction and refeeding with normal chow (CUFA rats); RT: CUFA rat supplemented with testosterone; ASST, Attentional set-shifting task; SD, Simple discrimination; CD, Compound discrimination; R1, First reversal; IDS, Intra-dimensional set-shifting; R2, Second reversal; EDS, Extradimensional set-shifting; R3, Third reversal; IRS-1, insulin receptor substrate 1, AKT, serine/threonine kinase 1; AR, androgen receptor; NR2A, N-methyl-d-aspartate receptors subtype 2A, NR2B, N-methyl-d-aspartate receptors subtype 2B

Article Snippet: Proteins extracts from the cerebral cortex of rats were prepared and immunoprecipitation was performed as reported previously.3 The antibodies used for immunoblot assays were anti-PKB (Protein kinase B)/Akt (1: 1000, Cell Signaling Technology), anti-phosphoPKB/Akt (Ser473) (1: 1000, Cell Signaling Technology), anti-NR2A (1:2000, Proteintech Group, Inc.), anti-NR2B (1:2000, Proteintech Group, Inc.), anti-AR (androgen receptor) (Proteintech Group, Inc), and anti-GAPDH (1:3000, Gene Tex).

Techniques: Expressing, Comparison, Phospho-proteomics

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: FCRL3, an autoimmune susceptibility gene, has inhibitory potential on B-cell receptor-mediated signaling.

doi: 10.4049/jimmunol.0901982

Figure Lengend Snippet: FIGURE 2. Coligation of FcRIIB/FCRL3 chimeric protein inhibits BCR-induced tyrosine phosphorylation and Ca2 influx in ST486 cells. A, ST486 transfectants of chimeric proteins were stimulated with F(ab)2 of anti-IgM (10.0 g/ml) or intact anti-IgM (15 g/ml) for 3 min before analysis of whole-cell phosphorylation of tyrosine residues detected by antiphosphotyrosine Ab (PY20). Blots were striped and reprobed using anti--actin Ab. The densities of phoshotyrosine proteins were quantified and shown on the top of figure, where each transfectant stimulated with F(ab)2 anti-IgM for 3 min is set to be 1.00. B, Whole lysates from A were immunoblotted against phosphor-Syk (Tyr352) and phosoho-PLC2 (Tyr1217). Blots were striped and reprobed with anti-Syk and PLC2 Ab, respectively. The densities of phosphor-Syk and phosoho-PLC2 were quantified and shown on the top of figure, where each transfectant stimulated with F(ab)2 anti-IgM is set to be 1.00. C, Whole lysates from A were immunoprecipitated with anti-HA Ab and immunoblotted with anti-phosphotyrosine Ab. Blots were striped and reprobed with anti-HA Ab. Data are representative of four separate experiments. D, ST486 transfectants of indicated chimeric proteins were loaded with fura-2 and stimulated with 10.0 g/ml F(ab)2 of anti-IgM (solid thick line) or 15 g/ml intact anti-IgM (dotted line). Chimeric proteins were also ligated alone with biotinylated 5.0 g/ml anti-murine FcRIIB (2.4G2) followed by 10.0 g/ml streptavidin (solid thin line). The results are representative of three independent experiments.

Article Snippet: Anti-hemagglutinin (HA) Ab (12CA5) was obtained from Roche Diagnostics; anti-phospho-Syk (Tyr352) and anti-phosphoPLC 2 (Tyr1217) Abs from Cell Signaling Technologies; biotinylated antimurine Fc RIIb (2.4G2) and HRP or PE-coupled anti-phosphotyrosine Abs (PY20) from BD Biosciences; anti-PLC 2, anti-SHP-1, anti-SHP-2, anti-SHIP and agarose-conjugated anti-HA Abs from Santa Cruz Biotechnology; anti-ZAP70 Ab from eBioscience, and intact and F(ab )2 of rabbit anti-human IgM and goat anti-chicken IgY from Jackson ImmunoResearch Laboratories.

Techniques: Phospho-proteomics, Transfection, Immunoprecipitation