Journal: International journal of medical sciences

Article Title: P66Shc is increased in peripheral blood mononuclear cells of the patients with obstructive sleep apnea.

doi: 10.7150/ijms.80343

Figure Lengend Snippet: Figure 2. P66Shc mRNA levels (n=73) were positively correlated with (A) plasma 3-NT, (B) oxLDL, and (C) AOPP levels by Spearman’s rank correlation coefficient.

Article Snippet: Enzyme-linked Immunosorbent Assay The level of plasma 3-NT (Elabscience, Catalog#E-EL-0040c, China), oxLDL (Elabscience, Catalog#E-EL-H6021, China) and AOPP (Abbkine, Catalog#KTB1060, China) were detected with the enzyme-linked immunosorbent assay (ELISA) commercial kits according to the recommendations of the manufacturers.

Techniques: Clinical Proteomics

Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: The Scavenger Receptor CD68 Regulates Platelet Mediated Oxidized Low-Density Lipoprotein (oxLDL) Deposition in Atherosclerotic Vessels at an Early Stage of Atherosclerosis in LDLR -/- /ApoBec -/- Mice.

doi: 10.33594/000000048

Figure Lengend Snippet: Fig. 1. (A) Schematic representation of the domain structure of Fc-CD68 fusion proteins. IgK: Ig Kappa leader sequence, hinge region, Fragments crystalizable (Fc) from human IgG1 or IgG2 were fused with the extracellular domain of CD68 using different linkers. L: short linker (GGR), Hel. L: linker with supposed rigid, helical conformation, Flex. L: linker with supposed flexible structure. (B) Coomassie-stained SDS-PAGE and Western-Blot analysis of the purified Fc-CD68 fusion proteins. A 4-20% PAA gradient gel was used for all analysis. A: Coomassie staining. B: Western Blot with an anti human Fc antibody. C: Western-Blot with an anti CD68 antibody. (C) Binding ELISA with lipoproteins oxLDL, HDL and LDL (1 µg/well) immobilized on a 96-well Maxisorp plate. Dilution series of the different Fc-fusion proteins in the soluble phase were tested for binding. The Means ± SEM of 4 independent experiments. (D) Binding ELISA with immobilized oxLDL. A dilution series of oxLDL (0.004-3 µg/well) was immobilized on a 96-well Maxisorp plate and binding of the different Fc-CD68 fusion proteins was tested using a fixed concentration in the soluble phase (10 µg/ ml). The Means ± SEM of 4 independent experiments. (E) Binding ELISA with immobilized fusion proteins. A dilution series of the Fc-CD68 fusion proteins (0,004 - 3 µg/well) was immobilized. oxLDL (10 µg/ml) in the soluble phase was tested for binding. The Means ± SEM of 4 independent experiments are shown.

Article Snippet: OxLDL content of the serum samples was measured using the ox-LDL/MDA Addukt ELISA kit (Immundiagnostik, #k 7810) or ELISA Kit for oxLDL for mice (cloud-clone Corp, Caty, Tx, USA) according to manufacturer’s protocol.

Techniques: Sequencing, Staining, SDS Page, Western Blot, Purification, Binding Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: bioRxiv

Article Title: Mast cell-driven adipocyte lipolysis promotes post-expansive VAT atrophy and ectopic hepatic steatosis

doi: 10.64898/2026.01.07.698076

Figure Lengend Snippet: ( a ) Heatmap of MC mediator-related gene expression in EAT in WT mice at week 0 and at week 10 of HFD feeding. n=3/ea. ( b ) Real-time PCR analysis of MC mediator-related genes Mcpt4 (encodes mouse mast cell protease 4 (mMCP-4), Cma1 (encodes mMCP-5), Hdc (encodes histidine decarboxylase), Ext1 (encodes exostosin-1), Ext2 , Ndst1 (encodes N-deacetylase/N-sulphotransferase 1), Ndst2 , Tph1 (encodes tryptophan hydroxylase 1), and Hpgds (encodes hematopoietic prostaglandin D synthase) in EAT in WT mice at week 0, 10, 20 of HFD feeding. n=6-8/ea. ( c/d ) Serum ( c ) and EAT ( d ) 5-HT levels in HFD-fed WT mice. n=6-8/ea. ( e ) EAT 5-HT levels in WT mice received i.p. injections of saline or DSCG from week 0 to 20 of HFD feeding. n=6-8/ea. ( f ) Real-time PCR analysis of Tph1 mRNA levels in adipocytes, SVFs, MC-depleted SVFs, and MCs from WT EAT in HFD-fed mice for 16 weeks. n=5-8/ea. ( g ) 5-HT + (red) and CD117 + (green), 5-HT + (red) and FceRI + (green) immunofluorescent double staining in WT EAT in HFD-fed mice for 16 weeks. ( h ) Correlations between serum LDL and 5-HT levels, IgE and 5-HT levels, LDL and NEFA levels, IgE and NEFA levels, and 5-HT and NEFA levels during EAT atrophy in HFD-fed mice. n=27. ( i ) Correlations between serum LDL and 5-HT levels, IgE and 5-HT levels, LDL and NEFA levels, IgE and NEFA levels, and 5-HT and NEFA levels in MASLD patients. n=50. ( j - l ) Differentiated 3T3-L1 adipocytes were treated with or without live BMMCs, BMMC lysates, LDL, LDL-activated BMMCs, DSCG- and LDL-treated BMMCs, or 5-HT. n=3/ea. ( j ) Representative images of oil-red O staining, media glycerol and NEFA levels in 3T3-L1 adipocytes. Scale bars: 50 µm. ( k/l ) Immunoblot analysis ( k ) and quantification of ( l ) ATGL, HSL and SIRT1 relative to β-actin, p-HSL(565) and p-HSL(563) relative to HSL, and p-AMPKα relative to AMPKα in 3T3-L1 adipocytes. Data are mean ± SEM. Pearson’s correlation test for h and i . P < 0.05 was considered significant correlation. Mann-Whitney U test for e . Kruskal-Wallis H test with Dunn’s post hoc adjustment for b-d , and f . Welch’s t-test for j and l . * p < 0.05, ** p < 0.01, *** p < 0.001. Different letters indicate statistically significant difference.

Article Snippet: To activate BMMCs, the cells were treated with 1 mg/mL LDL (Lee Biosolutions, Inc, Maryland Heights, MO Cat# 360-10).

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Histone Deacetylase Assay, Saline, Double Staining, Staining, Western Blot, MANN-WHITNEY

Journal: Reproduction

Article Title: Regulation of bovine oviductal NO synthesis by follicular steroids and prostaglandins

doi: 10.1530/rep-15-0254

Figure Lengend Snippet: Figure 3 (A) Cyclic changes of E-prostanoid receptor 2 (PTGER2), PTGER4, and F-prostanoid receptor (PTGFR) mRNA expressions in tissues collected from the ampulla and isthmus of the bovine oviduct (mean ± s.e.m., n = 5 oviducts). Significance of differences was assessed by the analysis of variance (ANOVA) followed by Tukey–Kramer test for multiple comparisons. There were no significant differences (P > 0.05). (B) Immunohistochemical analysis of PTGER and PTGFR protein distributions in the ampulla and isthmus of the oviduct obtained from cows at the day of ovulation (L: lumen, E: epithelium, ST: stroma, SM: smooth muscle). Green color (Alexa 488) indicates each target protein (PTGER2, PTGER4, or PTGFR), and blue color (DAPI) indicates nuclei of the cells. Scale is the same in all the photomicrographs.

Article Snippet: The sections were incubated with specific primary antibodies for NOS2 (160862; Cayman Chemical), NOS3 (160880; Cayman Chemical), ESR1 (SAB2100712; Sigma-Aldrich), PGR (SAB4502185; SigmaAldrich), PTGER2 (ab167171; Abcam), PTGER4 (ab133170; Abcam), or PTGFR (101802; Cayman Chemical) overnight at 4°C, washed with PBS three times, and incubated with secondary antibody for rabbit-IgG conjugated with Alexa 488 (21206; Life Technologies) for 1 h at room temperature, washed with PBS three times, covered with ProLong Gold Antifade Reproduction (2016) 151 577–587 www.reproduction-online.org Reagent with DAPI (36935; Life Technologies), and observed under a fluorescence microscope (FSX100; Olympus).

Techniques: Immunohistochemical staining

Journal: Reproduction

Article Title: Regulation of bovine oviductal NO synthesis by follicular steroids and prostaglandins

doi: 10.1530/rep-15-0254

Figure Lengend Snippet: Figure 5 (A) Effects of prostaglandin (PG) E2 and PGF2α on NOS2 mRNA expression in ampullary and isthmic oviductal epithelial cells (mean ± s.e.m., n = 5 oviducts). Significance of differences was assessed by the analysis of variance (ANOVA) followed by Tukey– Kramer test for multiple comparisons. Different superscript letters indicate significant differences (P < 0.05). (B) Effects of selective antagonists of each PTGER on NOS2 mRNA expression in isthmic oviductal epithelial cells (mean ± s.e.m., n = 7 oviducts). AH6809 (PTGER2 antagonist) or AH23848 (PTGER4 antagonist) was co-incubated with PGE2 (1 μmol/L) for 1 h. Significance of differences was assessed by the analysis of variance (ANOVA) followed by Tukey–Kramer test for multiple comparisons. Different superscript letters indicate significant differences (P < 0.05).

Article Snippet: The sections were incubated with specific primary antibodies for NOS2 (160862; Cayman Chemical), NOS3 (160880; Cayman Chemical), ESR1 (SAB2100712; Sigma-Aldrich), PGR (SAB4502185; SigmaAldrich), PTGER2 (ab167171; Abcam), PTGER4 (ab133170; Abcam), or PTGFR (101802; Cayman Chemical) overnight at 4°C, washed with PBS three times, and incubated with secondary antibody for rabbit-IgG conjugated with Alexa 488 (21206; Life Technologies) for 1 h at room temperature, washed with PBS three times, covered with ProLong Gold Antifade Reproduction (2016) 151 577–587 www.reproduction-online.org Reagent with DAPI (36935; Life Technologies), and observed under a fluorescence microscope (FSX100; Olympus).

Techniques: Expressing, Incubation

Journal: Reproduction

Article Title: Regulation of bovine oviductal NO synthesis by follicular steroids and prostaglandins

doi: 10.1530/rep-15-0254

Figure Lengend Snippet: Figure 7 Proposed model to illustrate region-specific regulations of nitric oxide (NO) synthesis in the ampulla and isthmus of the oviduct. Before ovulation, high level of estradiol-17β (E2) released from follicles systemically suppressed NO production by isthmic oviductal epithelial cells via estrogen receptor α (ESR1). Because NO stimulates the motility of spermatozoa (Miraglia et al. 2011), suppressing NO by E2 in the isthmus may contribute to decreasing the motility of spermatozoa and promoting their binding to epithelial cilia until they become hyperactivated. However, prostaglandin (PG), E2, and PGF2α derived from follicular fluid directly stimulate NO production by ampullary oviductal epithelial cells immediately after ovulation via E-prostanoid receptor type 2 (PTGER2) and F-prostanoid receptor (PTGFR) respectively. Because NO contributes to the survival of oocyte and embryos (Manser et al. 2004, Goud et al. 2005), PGs may play roles in maintaining the optimal concentrations of NO in the ampulla for the survival of oocyte and embryos.

Article Snippet: The sections were incubated with specific primary antibodies for NOS2 (160862; Cayman Chemical), NOS3 (160880; Cayman Chemical), ESR1 (SAB2100712; Sigma-Aldrich), PGR (SAB4502185; SigmaAldrich), PTGER2 (ab167171; Abcam), PTGER4 (ab133170; Abcam), or PTGFR (101802; Cayman Chemical) overnight at 4°C, washed with PBS three times, and incubated with secondary antibody for rabbit-IgG conjugated with Alexa 488 (21206; Life Technologies) for 1 h at room temperature, washed with PBS three times, covered with ProLong Gold Antifade Reproduction (2016) 151 577–587 www.reproduction-online.org Reagent with DAPI (36935; Life Technologies), and observed under a fluorescence microscope (FSX100; Olympus).

Techniques: Binding Assay, Derivative Assay