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: Cell Death Discovery

Article Title: MTA1 aggravates experimental colitis in mice by promoting transcription factor HIF1A and up-regulating AQP4 expression

doi: 10.1038/s41420-022-01052-y

Figure Lengend Snippet: A The statistical analysis for 30-day body weight of control mice and 2% DSS-treated mice. B Statistics for colon length of control mice and 2% DSS-induced mice. * p < 0.05 vs . the control group. C HE staining of cross-cut colon tissues in mice (200×). D Detection of CD45 positive cell rate in colon tissues of mice. The measurement data were expressed by mean ± standard deviation. Data between two groups were compared by unpaired student’s t test, and data at different time points were compared by repeated measures ANOVA, followed by Tukey’s post hoc tests. n = 8. *** p < 0.001.

Article Snippet: After conventional steps, the slices were then subjected to overnight incubation at 4 °C with rabbit anti-human against AQP4 (1:1000, ab90088, Abcam, Cambridge, UK) and CD45 (1:200, #13917, Cell Signaling Technologies [CST], Beverly, MA).

Techniques: Control, Staining, Standard Deviation

Journal: Cell Death Discovery

Article Title: MTA1 aggravates experimental colitis in mice by promoting transcription factor HIF1A and up-regulating AQP4 expression

doi: 10.1038/s41420-022-01052-y

Figure Lengend Snippet: A The statistic analysis for the changes in mouse weight after treatment with TGN-020. B The colon length of the mice after treatment with TGN-020. C HE staining to observe the pathological changes of colon tissues in TGN-020 treated mice (200×). D Statistical scoring results. E Graphical quantification of the colon wall thickness. F TUNEL staining analysis the apoptosis of colon epithelial cells in TGN-020 treated mice. G Immunohistochemistry of the expression of CD45 and AQP4. * p < 0.05. The measurement data were expressed as mean ± standard deviation. Data between two groups were compared by the unpaired student’s t test, and data among multiple groups were compared by one-way ANOVA, followed by Tukey’s post hoc tests. The experiment was conducted three times independently. n = 8; *** p < 0.001.

Article Snippet: After conventional steps, the slices were then subjected to overnight incubation at 4 °C with rabbit anti-human against AQP4 (1:1000, ab90088, Abcam, Cambridge, UK) and CD45 (1:200, #13917, Cell Signaling Technologies [CST], Beverly, MA).

Techniques: Staining, TUNEL Assay, Immunohistochemistry, Expressing, Standard Deviation