Journal: Aging and Disease

Article Title: IFNγ Transcribed by IRF1 in CD4 + Effector Memory T Cells Promotes Senescence-Associated Pulmonary Fibrosis

doi: 10.14336/AD.2023.0320

Figure Lengend Snippet: IFNγ produced by pulmonary CD4 + T EM cells promotes senescence and epithelial-to-mesenchymal transition of AT2 cells with aging. (A) Representative micrographs of pulmonary cells immunofluorescently stained for CD4, CD44, and IFNγ with DAPI staining the nucleus. (B) The percentage of CD4-, CD44-, and IFNγ-positive cells. **p < 0.01 compared with the 2-month-old group. Statistical analysis was performed with Student’s t-test. (C) Western blots showing SFTPC, α-SMA, TGF-β1, Smad2, p-Smad2(Ser465/467), p-Smad2/3(Ser423/425), Snail, IL-11, MEK1/2, p-MEK1/2 (Ser217/221), ERK1/2, p-ERK1/2(Thr202/Tyr204), and p16 in AT2 cells treated with different concentrations (0, 20, 40, 60, 80, and 100 ng/mL) of IFNγ for 48 h. β-actin was used as the loading control. (D) Protein levels relative to β-actin were assessed by densitometric analysis and normalized to the 0 ng/mL IFNγ-treated group. Three biological replicates were used per experiment (N = 3). Values are the mean ± SEM of six determinations. *p < 0.05, **p < 0.01, ***p < 0.001 compared with 0 ng/mL IFNγ-treated group. (E) Western blots showing SFTPC, α-SMA, TGF-β1, Smad2, p-Smad2(Ser465/467), p-Smad2/3(Ser423/425), Snail, IL-11, MEK1/2, p-MEK1/2(Ser217/221), ERK1/2, p-ERK1/2(Thr202/Tyr204), and p16 in AT2 cells treated with different groups of sorted cells, IFNγ, and anti-IFNγ. β-actin was used as the loading control. (F) Protein levels relative to β-actin were assessed by densitometric analysis and normalized to the untreated group. Three biological replicates were used per experiment (N = 3). Values are the mean ± SEM of three determinations. *p < 0.05, **p < 0.01, ***p < 0.001 compared with the untreated group; # p < 0.05, ## p < 0.01, ### p < 0.001 compared with the 2-month-old CD4 + T EM cell treated group; & p < 0.05, && p < 0.01, &&& p < 0.001 compared with the CD4 + T EM cell treated group. Statistical analysis was performed with one-way ANOVA.

Article Snippet: Primary antibodies against p19 (ab80, Abcam, USA), p21 (sc-471, Santa Cruz Biotechnology Inc., USA), p16 (ab211542, Abcam, USA), p53 (sc-126, Santa Cruz Biotechnology Inc., USA), SFTPC (ab211326, Abcam, USA), collagen 1 (#1310-08, Southern Biotech), α-SMA (ab5694, Abcam, USA), TGF-β1 (ab64715, Abcam, USA), Smad2 (sc-101153, Santa Cruz Biotechnology Inc., USA), phospho-Smad2 (Ser465/467) (#3108, Cell Signaling Technology, USA), Snail (#3879, Cell Signaling Technology, USA), IL-11 (sc-133063, Santa Cruz Biotechnology Inc., USA), IL-11Rα1 (sc-130920, Santa Cruz Biotechnology Inc., USA), MEK1/2 (sc-81504, Santa Cruz Biotechnology Inc., USA), phospho-MEK1/2 (sc-81503, Santa Cruz Biotechnology Inc., USA), ERK1/2 (#4695, Cell Signaling Technology, USA), pERK1/2 (Thr202/Tyr204) (#4370, Cell Signaling Technology, USA), phospho-Smad2/3 (Ser423/425) (sc-11769, Santa Cruz Biotechnology Inc., USA; #ab52903, abcam, USA), IRF1 (#11335-AP, Proteintech, USA), FLI1 (ab133485, Abcam, USA), IL-17A (sc-374218, Santa Cruz Biotechnology Inc., USA), and IFNγ (#15365, Proteintech, USA) were used. β-actin (BS6007M, Bioworld Technology, USA) was the loading control for the cytoplasmic fraction and total cell protein.

Techniques: Produced, Staining, Western Blot

Journal: Journal of Translational Medicine

Article Title: PTPRH promotes the progression of non-small cell lung cancer via glycolysis mediated by the PI3K/AKT/mTOR signaling pathway

doi: 10.1186/s12967-023-04703-5

Figure Lengend Snippet: PTPRH enhanced glycolysis via the phosphatidylinositol-3-kinase (PI3K)/Protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway. A GSEA to identify enriched signaling pathways for PTPRH. B Representative immunostaining images after staining of PI3K/AKT/mTOR signaling pathway-related proteins in A549 xenografts. C , D Relative expression levels of PI3K/AKT/mTOR signaling pathway-related proteins in the sh-PTPRH and sh-NC groups by western blotting. E Western blotting results showing the effects of sh-PTPRH and the PI3K activator 740Y-P alone and in combination on the expression levels of glycolysis-related proteins. F Western blotting results showing the effects of OE-PTPRH and the PI3K inhibitor LY294002 alone and in combination on the expression levels of glycolysis-related proteins

Article Snippet: Rabbit anti-PKM2 (#4053), rabbit anti-LDHA (#3582), rabbit anti-Akt (#9272), rabbit anti-phospho-Akt (Ser473) (#9271), rabbit anti-phospho-PI3K (Thr202/Tyr204) (#4370), rabbit anti-PI3K (#4257), rabbit anti-mTOR (#2983), rabbit anti-phospho-mTOR (#5536), and mouse anti-β-actin (#3700) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Immunostaining, Staining, Expressing, Western Blot

Journal: Journal of Translational Medicine

Article Title: PTPRH promotes the progression of non-small cell lung cancer via glycolysis mediated by the PI3K/AKT/mTOR signaling pathway

doi: 10.1186/s12967-023-04703-5

Figure Lengend Snippet: PTPRH promoted proliferation and invasion via PI3K/AKT/mTOR signaling pathway-mediated glycolysis. A , B EdU assay to verify the individual and combined effects of sh-PTPRH and the PI3K inhibitor LY294002 on the number of cells undergoing DNA replication. C , D Flow cytometry assays to detect the reversal of the proapoptotic effect of sh-PTPRH after treatment with the PI3K activator 740Y-P in H460 and A549 cells

Article Snippet: Rabbit anti-PKM2 (#4053), rabbit anti-LDHA (#3582), rabbit anti-Akt (#9272), rabbit anti-phospho-Akt (Ser473) (#9271), rabbit anti-phospho-PI3K (Thr202/Tyr204) (#4370), rabbit anti-PI3K (#4257), rabbit anti-mTOR (#2983), rabbit anti-phospho-mTOR (#5536), and mouse anti-β-actin (#3700) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: EdU Assay, Flow Cytometry

Journal: Journal of Translational Medicine

Article Title: PTPRH promotes the progression of non-small cell lung cancer via glycolysis mediated by the PI3K/AKT/mTOR signaling pathway

doi: 10.1186/s12967-023-04703-5

Figure Lengend Snippet: PTPRH increased invasion, FDG uptake and lactate levels via PI3K/AKT/mTOR signaling pathway-mediated glycolysis. A , B Transwell assays to detect the individual and combined effects of sh-PTPRH and the PI3K activator 740Y-P and OE-PTPRH and the PI3K inhibitor LY294002 on the invasion ability of H460 and A549 cells. C , D 18 F-FDG uptake assays to detect the individual and combined effects of sh-PTPRH and the PI3K activator 740Y-P and OE-PTPRH and the PI3K inhibitor LY294002 on the FDG uptake ability of H460 and A549 cells. E , F Cell metabolism assays to detect the individual and combined effects of sh-PTPRH and the PI3K activator 740Y-P and OE-PTPRH and the PI3K inhibitor LY294002 on the lactate levels of H460 and A549 cells. G A diagram showing the possible mechanism underlying the modulation of glycolysis by PTPRH

Article Snippet: Rabbit anti-PKM2 (#4053), rabbit anti-LDHA (#3582), rabbit anti-Akt (#9272), rabbit anti-phospho-Akt (Ser473) (#9271), rabbit anti-phospho-PI3K (Thr202/Tyr204) (#4370), rabbit anti-PI3K (#4257), rabbit anti-mTOR (#2983), rabbit anti-phospho-mTOR (#5536), and mouse anti-β-actin (#3700) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: