Fig 1: Immunohistochemical analysis for Fas. Representative images of Fas detection in subcutaneous abdominal fat from obese patient with pre-diabetic condition and obese patient with normo-glycemic condition. Immunoistochemistry of obese patient with pre-diabetic condition in metformin therapy (group 1, left upper part, A) vs. obese patient with pre-diabetic condition in placebo therapy (group 2, right upper part, B) vs. obese patient with normo glycemic condition (group 3, in the left inferior part, C); In the right inferior part (D), expression of FAS in obese patient with pre-diabetic condition in metformin therapy (group 1, red color), obese patient with pre-diabetic condition in placebo therapy (group 2, gray color), and in obese patient with normo glycemic condition (group 3, green color). Results are mean ± SD of 5 independent experiments. Results are mean ± SD of 5 independent experiments. Group 1 vs. group 3, *P <0.05; group 2 vs. group 3, **p <0.05.
Fig 2: SOCS3 prevents AA induced by transfer of CD8+ T cells through inhibiting IFN-γ signalingCD8+ T cells isolated from SDLNs of AA-affected mice were subcutaneously injected into normal C3H/HeJ recipients. Mice then were subcutaneously injected beginning the day of injection with 250 μg of SOCS3 or IgG (n = 9 mice/group). (A) The onset of alopecia was inhibited by treatment of SOCS3. (B) Time course of onset of AA in SOCS3–treated mice and control mice was shown as weeks after CD8+ T cells injection. (C) CD8+ T cells were isolated from skins of SOCS3-treated C3H/HeJ AA mice and were used to assay the percentage of IFN-γ-producing CD8+ T cells. Data are representative of three independent experiments. (D) Percentage of IFN-γ-producing CD8+ T cells was quantitated by flow cytometric analysis. (E) CD8+ T cells were isolated from SDLNs of SOCS3-treated C3H/HeJ AA mice and were used to assay the level of CD44 and CD62L. SOCS3 treatment markedly decreases CD44high CD62Llow effector memory CD8+ T cells. Data are representative of three independent experiments. (F) Percentage of CD44high CD62Llow effector memory CD8+ T cells was quantitated by flow cytometric analysis. (G) IFN-γ mRNA level in the skin of injected recipients was assayed after treatment with SOCS3 or IgG control (n = 7 mice/group). (H) The Fas and MHC I mRNA level in the skin of injected recipients was assayed after treatment with SOCS3 or IgG control (n = 7 mice/group). *p < 0.05.
Fig 3: Effect of SCG on protein expression of TRAIL, DR5, FASL, FAS, IL-33, and caspase-3 in Con A-treated mice. (A) The expression of TRAIL. (B) The expression of DR5. (C) The expression of IL-33. (D) The expression of FASL. (E) The expression of FAS. (F) The expression of cleaved caspase-3. Data were expressed as mean ± SD (n = 3). ∗∗P < 0.01 compared with normal group; #P < 0.05, ##P < 0.01 compared with model group.
Fig 4: XIAP silence enhances dyslipidemia in PA-treated cells in vitro. (A) AML-12 and primary hepatocytes were transfected with XIAP specific siRNA sequence for 24 h, followed by transfection efficacy calculation using qPCR and western blot analysis. AML-12 and primary hepatocytes were subjected to XIAP siRNA transfection for 24 h, and then cells were incubated with 250 μM of PA for additional 24 h, followed by further investigation. (B) Representative western blot and qPCR analysis of the expression of ACCα, FAS and SCD1 in PA-induced hepatocytes and AML-12 cell line. (C) Determination of cellular TG and TC levels in AML-12 and hepatocytes was next detected. (D) qPCR analysis for genes associated with fatty acid metabolism, including CD36, FATP1, FABP1, CPT-1α and PPARα, in cells treated as indicated. For all bar plots shown, data are expressed as the mean ± SEM. n = 8 per group. ap < 0.05 vs. siXIAP or siRNA/PA.
Fig 5: Widespread changes of gene expressions and critical pathways in human lung adenocarcinoma cells A549 with GMDS knockdown. a Heatmap containing 739 differentially expressed genes in human lung adenocarcinoma cell line A549 infected with lentivirus expressing either Scr-shRNA (purple) or GMDS-shRNA (red) with the criteria P < 0.05 and ▏fold change ▏ > 1.5. Genes and samples were listed in rows and columns, respectively. A colour standard for normalized expression data was shown at the bottom of the microarray heatmap (green represents downregulated genes while red represents upregulated genes). b Gene expression changes identified in microarray were confirmed using real-time quantitative PCR for selected genes CASP8, MAP3K7, CDKN1A, FAS, JUN, DDIT3, VEGFA, SKA1 and MAD2L1 in human lung adenocarcinoma cell line A549 infected with lentivirus expressing either Scr-shRNA or GMDS-shRNA. Histogram shown here was one out of three independent experiments (p < 0.01) and normalized to GAPDH. c Protein level of FAS, VEGFA, DDIT3, JUN and CDKN1A in human human lung adenocarcinoma cell line A549 infected with lentivirus expressing either Scr-shRNA or GMDS-shRNA. GAPDH was used as internal control. d Caspase3/7 activity analysis in A549 cells and H1299 cells infected with lentivirus expressing either Scr-shRNA or GMDS-shRNA. Data shown are the mean ± SEM of caspase3/7 activity from three independent experiments (**, p < 0.01)
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