Journal: Brain and behavior

Article Title: The behavioral and physiological effects of high-fat diet and alcohol consumption: Sex differences in C57BL6/J mice.

doi: 10.1002/brb3.708

Figure Lengend Snippet: FIGURE 6 Whole-brain BDNF. When normalized to their respective sex controls, males overall exhibited small but significant decreases to BDNF compared to females. Black bars indicate water, and gray bars indicate 10% EtOH. The dashed line indicates 100% RC H2O expression. ‡ significant sex difference at p < .05

Article Snippet: A low target concentration (working dilution 1:2) of sample and sample diluent buffer was created and then used in a BDNF ELISA (Mouse BDNF PicoKine ELISA Kit, Boster Biological Technology Co., Pleasanton, CA, USA).

Techniques: Expressing

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Magnesium isoglycyrrhizinate ameliorates radiation-induced pulmonary fibrosis by inhibiting fibroblast differentiation via the p38MAPK/Akt/Nox4 pathway.

doi: 10.1016/j.biopha.2019.108955

Figure Lengend Snippet: Fig. 4. MgIG regulated the expression of TGF-β1 and Nox4, and the phosphorylation of p38MAPK and Akt in vivo. The protein level of TGF-β1 in lung tissues at 12 weeks post-irradiation was determined using (A) im- munohistochemical staining (×200, n = 6) and (B) western blotting (n = 3), and quanti- tative analyses were performed. (C) TGF-β1 content in serum at 12 weeks post-irradiation was measured using an ELISA kit (n = 6). The protein levels of Nox4, and the phosphoryla- tion of p38MAPK and Akt in lung tissues at 12 weeks post-irradiation were observed using (D) immunohistochemical staining (×200, n = 6) and (E) western blotting (n = 3), and quanti- tative analyses were performed. All data were expressed as the mean ± SEM. #P < 0.05 vs. the control group; *P < 0.05 vs. the IR group; &P < 0.05 vs. the IR + MgIG group; Bar =50 μm.

Article Snippet: The serum was used to measure the TGF-β1 concentration using an ELISA kit (EK0515, Boster Bioengineering Institute, Huhan, China), according to the manufacturer's instructions.

Techniques: Expressing, Phospho-proteomics, In Vivo, Irradiation, Staining, Western Blot, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Control

Journal: Nature communications

Article Title: VSIG4 inhibits proinflammatory macrophage activation by reprogramming mitochondrial pyruvate metabolism.

doi: 10.1038/s41467-017-01327-4

Figure Lengend Snippet: Fig. 1 Vsig4−/−mice are more susceptible to HFD-induced obesity with insulin resistance. Eight-week-old male Vsig4−/−mice and age-matched C57BL/6 WT controls were fed a HFD. a Body weight was measured and compared. The obese mice were sacrificed after 10 weeks of HFD feeding. b Fat distribution was detected by μCT. Yellow indicates subcutaneous fat and brown indicates that visceral fat. c Measurement of abdominal wall fat, perirenal fat, serum triglyceride, cholesterol, and free fatty acid. d Representative liver H&E staining (left), and intrahepatic triglyceride contents (right), scale bar = 20 μm, n = 10 per group. e Representative the architecture of adipose tissues stained by H&E (left), adipocyte size and cell numbers was calculated (right), scale bar = 20 μm, n = 10 per group. f The 15-h-fasting blood glucose levels and 5-h-fasting serum insulin levels. g GTT and ITT were performed in theses obese mice, n = 6 per group. h Western blot of the AKT, p-Aktser473, and p-IRS-1 in VAT, muscle and liver tissues of obese mice after 4 min of insulin administration, n = 4 per group. ATMs were isolated from obese mice. i Flow cytometry analyzing pro-IL-1β, IFN-γ, and TNF. Cytokines in VAT were detected by j qRT-PCR and k western blot. Error bar, s.e.m. *p < 0.05, **p < 0.01 and ***p < 0.001 (Student’s t-test). Data are representative of five (a) and three (c–i) independent experiments

Article Snippet: ELISA Kits, including TNF (#EK0527), IL-6 (#EK0411), IL-1β (#EK0394), and IL-12 p40 (#EK0932) were from Boster Ltd. (Wuhan, China).

Techniques: Staining, Western Blot, Isolation, Flow Cytometry, Quantitative RT-PCR

Journal: Nature communications

Article Title: VSIG4 inhibits proinflammatory macrophage activation by reprogramming mitochondrial pyruvate metabolism.

doi: 10.1038/s41467-017-01327-4

Figure Lengend Snippet: Fig. 2 Vsig4 deficiency exacerbates MHV-3-induced fulminant hepatitis. The Vsig4−/−mice and age-matched C57BL/6 WT littermates were infected with MHV-3 (100 PFU/mouse) via i.p. injection. a The survival was monitored. b H&E staining of liver, and TUNEL staining of cell apoptosis, scale bar = 20 μm, n = 5–8 per group, arrow indicated positive cells. c Serum ALT and AST levels at 0 h and 48 h post infection (PI), n = 5–8 per group. d Plaque assay of virus titers in livers at 48 h PI. e qRT-PCR analyzing proinflammatory cytokines in PEMs at 12 h and in liver tissues at 72 h of MHV-3 infection. f Flow cytometry analyzing TNF, pro-IL1-β, and IL-6 from PEMs after 12 h of virus infection. g Western blot analyzing proinflammatory cytokines in infected livers at 24 h and 48 h PI, n = 4 per group. h ELISA of serum concentration of proinflammatory mediators, n = 5–10 per group. Error bar, s.e.m. *p < 0.05, **p < 0.01, ***p < 0.001 and NS, p > 0.05. a was analyzed by log-rank test and others are calculated by Student’s t-test. Data are representative of six (a) and three (b–f, h) independent experiments

Article Snippet: ELISA Kits, including TNF (#EK0527), IL-6 (#EK0411), IL-1β (#EK0394), and IL-12 p40 (#EK0932) were from Boster Ltd. (Wuhan, China).

Techniques: Infection, Injection, Staining, TUNEL Assay, Plaque Assay, Virus, Quantitative RT-PCR, Flow Cytometry, Western Blot, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Nature communications

Article Title: VSIG4 inhibits proinflammatory macrophage activation by reprogramming mitochondrial pyruvate metabolism.

doi: 10.1038/s41467-017-01327-4

Figure Lengend Snippet: Fig. 3 VSIG4 impedes LPS-induced macrophage M1 polarization in vitro. PEMs were treated with LPS (2 μg/ml), a qRT-PCR analysis of Il-1β and Tnf transcripts. b ELISA of cytokines in cultured supernatants. c Western blot analyzing cytokine protein expression. d Flow cytometry analyzing surface expression of activation markers. RAW264.7 cells stably infected with lentiviral control vectors (Len-cont.) or vectors encoding Vsig4 (Len-Vsig4), cells were further treated with LPS (2 μg/ml), e qRT-PCR analysis of Il-1β, Il-6, and Tnf transcripts. f ELISA detecting cytokines in cultured supernatant. g Flow cytometry analyzing surface expression of CD40. h C3−/−BMDMs were tranfected to overexpress VSIG4, and cells were further treated with LPS (2 μg/ ml), the secretion of IL-6 and IL-1β was detected by ELISA. Error bar, s.e.m. *p < 0.05, **p < 0.01, ***p < 0.001 and NS, p > 0.05 (Student’s t-test). Data are representative of three independent experiments

Article Snippet: ELISA Kits, including TNF (#EK0527), IL-6 (#EK0411), IL-1β (#EK0394), and IL-12 p40 (#EK0932) were from Boster Ltd. (Wuhan, China).

Techniques: In Vitro, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Cell Culture, Western Blot, Expressing, Flow Cytometry, Activation Assay, Stable Transfection, Infection, Control

Journal: Nature communications

Article Title: VSIG4 inhibits proinflammatory macrophage activation by reprogramming mitochondrial pyruvate metabolism.

doi: 10.1038/s41467-017-01327-4

Figure Lengend Snippet: Fig. 5 VSIG4 triggers PDK2 expression in macrophages. Macrophages from WT and Vsig4−/−mice were collected. a qRT-PCR detection of 4 Pdk isoforms in BMDMs. b Western blot analyzing PDK2, p-PDH-E1αs300, p-PDH-E1αs293, and total PDH. c The location of p-PDH-E1αs300 in mitochondria was analyzed by immunofluoresence double staining, scale bar = 20μm. d Western blot of PDK2, p-PDH-E1αs300, p-PDH-E1αs293 in liver tissues at 0 h and 48 h PI. RAW264.7 cells were transfected to expression of Vsig4, and cells were further treated with LPS (2 μg/ml), e Western blot analysis of PDK2 and p-PDH- E1αs300. f PDH activity analysis, n = 6 per group. The expression of Pdk2 in RAW264.7 cells was silenced by shRNA or enhancing Pdk2 expression by lentivirus infection. g Seahorse analysis of OCR after 2 h of LPS treatment (up), and basal and maximal OCR of the indicated conditions was plotted in bar graphs (down), n = 5 per group. h Flow cytometric assay of mtROS secretion after LPS administration. i ELISA of IL-6 and TNF in cultured supernatants, n = 4 per group. j Flow cytometric assay of LPS-caused CD40 expression at 6 h. Error bar, s.e.m. *p < 0.05,**p < 0.01, ***p < 0.001 and NS, p > 0.05 (Student’s t-test). Data are representative of three independent experiments

Article Snippet: ELISA Kits, including TNF (#EK0527), IL-6 (#EK0411), IL-1β (#EK0394), and IL-12 p40 (#EK0932) were from Boster Ltd. (Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Double Staining, Transfection, Activity Assay, shRNA, Infection, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Nature communications

Article Title: VSIG4 inhibits proinflammatory macrophage activation by reprogramming mitochondrial pyruvate metabolism.

doi: 10.1038/s41467-017-01327-4

Figure Lengend Snippet: Fig. 8 Forced overexpression of Vsig4 improves MHV-3-induced hepatitis. C57BL/6 WT mice were infected with lentivirus (107 PFU/mouse) to induce the expression of Vsig4 in vivo, these mice were further infected with MHV-3 at day 6. a Liver Vsig4 gene transcription was analyzed by qRT-PCR at day 6, n = 5 per group. b Western blotting for PDK2, p-PDH-E1αs300, FGL2, and proinflammatory cytokines TNF, IL-6 and IL-1β in liver tissues at 72 h of MHV-3 infection, n = 4 per group. c The architecture of the liver tissues at 72 h of infection was compared by H&E staining, scale bar = 20 μm, n = 5 per group. d The survival was monitored for a total of 20 days. Error bar, s.e.m. a *p < 0.05 was analyzed by Student’s t-test, and d was analyzed by log-rank test. Data are representative of three independent experiments

Article Snippet: ELISA Kits, including TNF (#EK0527), IL-6 (#EK0411), IL-1β (#EK0394), and IL-12 p40 (#EK0932) were from Boster Ltd. (Wuhan, China).

Techniques: Over Expression, Infection, Expressing, In Vivo, Quantitative RT-PCR, Western Blot, Staining

Journal: Bioactive Materials

Article Title: Acellular nerve xenografts based on supercritical extraction technology for repairing long-distance sciatic nerve defects in rats

doi: 10.1016/j.bioactmat.2022.03.014

Figure Lengend Snippet: Evaluation of the cytotoxicity of the ANXs scaffold in vitro. (A, D) Living/dead double staining of Schwann cells grown on the ANXs scaffold for 3 days and 7 days (live: green, dead: red). (B, E) SEM images of Schwann cells growing on CD SD and CD + scCO 2 NG scaffolds for 7 days (the picture on the right is an enlarged view of the yellow area in the picture on the left). (C, F) Immunofluorescence images of Schwann cells growing on CD SD and CD + scCO 2 NG scaffolds for 7 days, respectively (S100: red, nucleus: blue). (G) Quantification of the number of live/dead double-stained Schwann cells in each region (0.36 mm 2 ). Data are presented as the mean ± SD (n = 3). (H) The CCK-8 assay was performed after 1, 3, 5 and 7 days of cell culture. Data are presented as the mean ± SD (n = 5). (I, J) Quantitative analysis of the GDNF and NGF expression levels of Schwann cells on the ANXs scaffold. Data are presented as the mean ± SD (n = 5). Statistical analysis: n.s. no significances, **p < 0.01, *p < 0.05.

Article Snippet: In brief, the medium of each group was centrifuged at 1500 rpm and 4 °C for 10 min, the concentration of NGF and BDNF in the supernatant was assessed using ELISA kits, the rat GDNF ELISA kit (EK0363, BOSTER, China) and the rat NGF/NGFβ ELISA kit (EK0471, BOSTER, China), and the absorbance of each well at 450 nm was determined using a spectrophotometer (EPOCH TAKE 3, Bio-Tek, USA).

Techniques: In Vitro, Double Staining, Immunofluorescence, Staining, CCK-8 Assay, Cell Culture, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Single‐cell RNA‐seq reveals altered NK cell subsets and reduced levels of cytotoxic molecules in patients with ankylosing spondylitis

doi: 10.1111/jcmm.17159

Figure Lengend Snippet: Expression of cytolytic molecules in plasma of HCs ( n = 16) and AS patients ( n = 16). The plasma levels of granzyme A (A), granzyme B (B) and granulysin (C) were determined by enzyme‐linked immunosorbent assay. Pearson correlation analysis was performed between granzyme A (D), granzyme B (E), granulysin (F) and disease activity (BASDAI). Horizontal lines and error bars show the mean ± SEM. ** p < 0.01; *** p < 0.001; ns = not significant

Article Snippet: Plasma levels of cytotoxic granules, including GZMA, GZMB and granulysin, were quantified using ELISA kits (#EK1162, #EK1114 and # EK1280, BOSTER Biological Technology).

Techniques: Expressing, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Activity Assay