serum sp d cat msfpd0 concentrations  (R&D Systems)

Journal: Frontiers in Pharmacology

Article Title: Neoshirakia japonica (Siebold & Zucc.) Esser [Euphorbiaceae] fruit suppresses obesity and obesity-induced inflammation in adipocytes, macrophages, and obese mice

doi: 10.3389/fphar.2025.1647343

Figure Lengend Snippet: Effects of NJFE on adipose tissue inflammation in high-fat diet (HFD)-induced obese mice. (A) F4/80 immunohistochemistry images of epididymal white adipose tissue (eWAT). Images were taken at 400× magnification. The scale bar represents 100 μm. The mRNA levels of two genes in eWAT, measured using RT-qPCR: (B) F4/80 and (C) Tnfα . (D) Expression levels of two adipogenesis-related genes in eWAT at the mRNA level. (E) Protein expression levels of adipogenesis-related proteins, PPARγ and C/EBPα, and phosphorylated NF-κB in eWAT, measured via Western blot analysis. The numbers on the right indicate the molecular weights of the protein bands (kDa). (F) A graph of PPARγ protein expression quantified based on Western blot band intensities. (G) Graph of C/EBPα protein expression quantified based on Western blot band intensities. (H) Graph of NF-κB phosphorylation levels quantified based on Western blot band intensities. The phosphorylation level was calculated by dividing the p-NF-κB band intensity by the NF-κB band intensity. All Western blot band intensities and RT-qPCR results were normalized using β-actin and are presented as fold changes relative to the expression levels in the control (HFD) group. All data are presented as means ± standard deviations. Statistical significance is denoted using symbols: “#” indicates a statistically significant difference ( p < 0.05) between the control groups, ND and HFD, and “**” indicates a p -value <0.01 for comparisons between the NJFE-treated and HFD mice.

Article Snippet: The assay was performed according to the manufacturer’s protocol using the Mouse TNFα ELISA Kit–Quantikine (R&D Systems, Oxon, United Kingdom).

Techniques: Immunohistochemistry, Quantitative RT-PCR, Expressing, Western Blot, Phospho-proteomics, Control

Journal: Frontiers in Pharmacology

Article Title: Neoshirakia japonica (Siebold & Zucc.) Esser [Euphorbiaceae] fruit suppresses obesity and obesity-induced inflammation in adipocytes, macrophages, and obese mice

doi: 10.3389/fphar.2025.1647343

Figure Lengend Snippet: The anti-inflammatory effect of NJFE on macrophage-mediated inflammatory responses. (A) Cell viability of RAW264.7 cells after NJFE treatment, as assessed using the CCK-8 assay. For the control (0), the same volume of DMSO as that used in the 32 μg/mL NJFE group was applied. (B) The concentrations (μM) of nitric oxide (NO) secreted by RAW264.7 macrophages in the cell culture medium. (C) The concentrations of TNFα secreted by RAW264.7 macrophages in the cell culture medium. The values are presented as fold change relative to the control (LPS). (D) The relative mRNA levels of Tnfα and Il-6 in RAW264.7 macrophages, measured using RT-qPCR. (E) A Western blot membrane showing the protein expression levels of iNOS and COX2. The numbers on the right indicate the molecular weights of the protein bands (kDa). (F) A graph of iNOS and COX2 protein expression levels quantified based on Western blot band intensities. Western blot band intensities and RT-qPCR results were normalized using β-actin and are presented as fold changes relative to the expression levels in the control (LPS) group. All data are presented as means ± standard deviations. Statistical significance is denoted using symbols: “#” indicates a statistically significant difference ( p < 0.05) between the control groups, CON and LPS, and “*” and “**” indicate p -values of <0.05 and <0.01 for comparisons between the NJFE-treated and LPS cells.

Article Snippet: The assay was performed according to the manufacturer’s protocol using the Mouse TNFα ELISA Kit–Quantikine (R&D Systems, Oxon, United Kingdom).

Techniques: CCK-8 Assay, Control, Cell Culture, Quantitative RT-PCR, Western Blot, Membrane, Expressing

Journal: Frontiers in Pharmacology

Article Title: Neoshirakia japonica (Siebold & Zucc.) Esser [Euphorbiaceae] fruit suppresses obesity and obesity-induced inflammation in adipocytes, macrophages, and obese mice

doi: 10.3389/fphar.2025.1647343

Figure Lengend Snippet: The alleviative effects of NJFE on adipocyte inflammation induced by lipogenesis and an activated macrophage-conditioned medium. (A) The relative mRNA levels of Tnfα and Il-6 in 3T3-L1 adipocytes. (B) A schematic detailing the treatment of 3T3-L1 cells with a RAW264.7 macrophage-conditioned medium (RAW-CM) in cell culture experiments. (C) A Western blot membrane showing the phosphorylation level of NF-κB and adiponectin protein expression. The numbers on the left indicate the molecular weights of the protein bands (kDa). (D and E) Graphs of (D) NF-κB phosphorylation levels and (E) adiponectin expression levels, both quantified based on Western blot band intensities. The relative mRNA levels, based on RT-qPCR analysis, of (F) Il-6 , (G) Tnfα , and (H) Mcp1 in RAW-CM-treated 3T3-L1 adipocytes. Western blot band intensities and RT-qPCR results were normalized using β-actin and are presented as fold changes relative to line 2. All data are presented as means ± standard deviations. Statistical significance is denoted using symbols: “#” indicates a statistically significant difference ( p < 0.05) between the control groups, lines 1 and 2, and “*” and “**” indicate p -values of <0.05 and <0.01 for compared to line 2.

Article Snippet: The assay was performed according to the manufacturer’s protocol using the Mouse TNFα ELISA Kit–Quantikine (R&D Systems, Oxon, United Kingdom).

Techniques: Cell Culture, Western Blot, Membrane, Phospho-proteomics, Expressing, Quantitative RT-PCR, Control