Journal: Military Medical Research

Article Title: High-salt-driven gut microbiota dysfunction aggravates prostatitis by promoting AHR/SGK1/FOXO1 axis-mediated Th17 cell differentiation

doi: 10.1186/s40779-025-00607-1

Figure Lengend Snippet: HSD promoted Th17 cell development and worsened prostate inflammation in EAP mice. a Simple schematic diagram of the implementation plan of the experiment. b HE staining and inflammation scores of prostate tissues (scale bar = 100 μm, n = 4). c Tactile allodynia development in the 4 groups of mice ( n = 4). d The levels of IL-1β, TNF-α, IL-17A, and IFN-γ in the serum of the mice in the 4 groups ( n = 4). e Proportion of Th1 and Th17 cells of CD4 + T cells detected by flow cytometry in the splenic lymphocytes of vaccinated mice from those 4 groups ( n = 4). f The infiltration of Th17 cells in prostate tissues from mice was determined by immunofluorescence (white arrowheads, scale bar = 100 μm, n = 4). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns non-significant. BF breeding feed, EAP experimental autoimmune prostatitis, HSD high-salt diet, NSD normal-salt diet, IL-1β interleukin-1β, TNF-α tumor necrosis factor-α, IFN-γ interferon-γ, IL-17A interleukin-17A, DAPI 4',6-diamidino-2-phenylindole, Th T helper

Article Snippet: The following reagents and antibodies were used: CFA (#F5881, Sigma, USA); commercial enzyme-linked immunosorbent assay (ELISA) kits for IFN-γ (#E-EL-M0048c), IL-1β (#E-EL-M0037), IL-17A (#E-EL-M0047c), tumor necrosis factor-α (TNF-α, #E-EL-M3063), and 5-HIAA (#E-EL-0075c) (Elabscience, China); anti-mouse CD4-fluorescein isothiocyanate (FITC, #553047), CD3e-allophycocyanin (APC, #561826), IFN-γ-phycoerythrin (PE, #562020), IL-17-PE (#559502), anti-human CD8-FITC (#555634), CD3-APC (#555342), IL-17-PE (#560436) (BD, USA); anti-CD3e (#BE0001-1), anti-CD28 (#BE0015-1), anti-IFN-γ (#BE0055), anti-IL-4 (#BE0045) (Bio X Cell, USA); IL-23 (#CS31), transforming growth factor-β1 (TGF-β1, #CA59), IL-6 (#CG39) (Novoprotein, China); anti-AHR (1:1000, #NB100-128, Novus, USA); anti-SGK1 (1:1000, #AF6789), anti-FOXO1 (1:1000, #AF6416), anti-p-FOXO1 (1:1000, #AF3417), anti-retinoic acid receptor-related orphan receptor gamma-t (RORγt; 1:1000, #DF3196), and anti-GAPDH (1:5000, #AF7021) (Affinity, USA).

Techniques: Staining, Flow Cytometry, Immunofluorescence

Journal: Military Medical Research

Article Title: High-salt-driven gut microbiota dysfunction aggravates prostatitis by promoting AHR/SGK1/FOXO1 axis-mediated Th17 cell differentiation

doi: 10.1186/s40779-025-00607-1

Figure Lengend Snippet: Transplantation of fecal microbiota from EAP mice with HSD elicited more severe prostate inflammation in psuedogerm-free mice. a Simple schematic diagram of the experimental workflow. b HE staining and inflammation scores of prostate tissues (scale bar = 100 μm, n = 4). c Development of tactile allodynia in mice belonging to the 4 groups ( n = 4). d The concentrations of IL-1β, TNF-α, IL-17A, and 5-HIAA in the serum of NOD mice in the 4 groups were measured ( n = 4). e Proportion of Th17 of CD4 + T cells, as determined by flow cytometry, among the splenic lymphocytes of immunized mice in the 4 groups ( n = 4). f The infiltration of Th17 cells in prostate tissues from mice was determined by immunofluorescence (white arrowheads, scale bar = 100 μm, n = 4). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns non-significant. NOD nonobese male diabetic/LtJ, EAP experimental autoimmune prostatitis, HSD high-salt diet, NSD normal-salt diet, FMT fecal microbiota transplantation, HE hematoxylin–eosin, IL-1β interleukin-1β, TNF-α tumor necrosis factor-α, IL-17A interleukin-17A, 5-HIAA 5-hydroxyindole acetic acid, DAPI 4',6-diamidino-2-phenylindole

Article Snippet: The following reagents and antibodies were used: CFA (#F5881, Sigma, USA); commercial enzyme-linked immunosorbent assay (ELISA) kits for IFN-γ (#E-EL-M0048c), IL-1β (#E-EL-M0037), IL-17A (#E-EL-M0047c), tumor necrosis factor-α (TNF-α, #E-EL-M3063), and 5-HIAA (#E-EL-0075c) (Elabscience, China); anti-mouse CD4-fluorescein isothiocyanate (FITC, #553047), CD3e-allophycocyanin (APC, #561826), IFN-γ-phycoerythrin (PE, #562020), IL-17-PE (#559502), anti-human CD8-FITC (#555634), CD3-APC (#555342), IL-17-PE (#560436) (BD, USA); anti-CD3e (#BE0001-1), anti-CD28 (#BE0015-1), anti-IFN-γ (#BE0055), anti-IL-4 (#BE0045) (Bio X Cell, USA); IL-23 (#CS31), transforming growth factor-β1 (TGF-β1, #CA59), IL-6 (#CG39) (Novoprotein, China); anti-AHR (1:1000, #NB100-128, Novus, USA); anti-SGK1 (1:1000, #AF6789), anti-FOXO1 (1:1000, #AF6416), anti-p-FOXO1 (1:1000, #AF3417), anti-retinoic acid receptor-related orphan receptor gamma-t (RORγt; 1:1000, #DF3196), and anti-GAPDH (1:5000, #AF7021) (Affinity, USA).

Techniques: Transplantation Assay, Staining, Flow Cytometry, Immunofluorescence

Journal: Military Medical Research

Article Title: High-salt-driven gut microbiota dysfunction aggravates prostatitis by promoting AHR/SGK1/FOXO1 axis-mediated Th17 cell differentiation

doi: 10.1186/s40779-025-00607-1

Figure Lengend Snippet: 5-HIAA supplementation ameliorated the symptoms of EAP caused by HSD. a The levels of 5-HIAA secreted into the serum of the mice were measured in 4 groups: Ctrl + NSD, Ctrl + HSD, EAP + NSD, and EAP + HSD groups ( n = 4). b Simple schematic diagram of the experimental workflow. c HE staining and inflammation scores of prostate tissues (scale bar = 100 μm, n = 4). d Tactile allodynia development in NOD mice in the 4 groups ( n = 4). e Secretion levels of IL-1β, TNF-α, and IL-17A in the serum of mice in the 5 groups ( n = 4). f Flow cytometry was used to determine the proportion of Th17 of CD4 + T cells among the splenic lymphocytes of immunized mice in the 5 groups ( n = 4). g The infiltration of Th17 cells in prostate tissues from mice was determined by immunofluorescence (white arrowheads, scale bar = 100 μm, n = 4). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns non-significant. NOD nonobese male diabetic/LtJ, BF breeding feed, EAP experimental autoimmune prostatitis, HSD high-salt diet, NSD normal-salt diet, HE hematoxylin–eosin, IL-1β interleukin-1β, TNF-α tumor necrosis factor-α, IL-17A interleukin-17A, 5-HIAA 5-hydroxyindole acetic acid, DAPI 4',6-diamidino-2-phenylindole

Article Snippet: The following reagents and antibodies were used: CFA (#F5881, Sigma, USA); commercial enzyme-linked immunosorbent assay (ELISA) kits for IFN-γ (#E-EL-M0048c), IL-1β (#E-EL-M0037), IL-17A (#E-EL-M0047c), tumor necrosis factor-α (TNF-α, #E-EL-M3063), and 5-HIAA (#E-EL-0075c) (Elabscience, China); anti-mouse CD4-fluorescein isothiocyanate (FITC, #553047), CD3e-allophycocyanin (APC, #561826), IFN-γ-phycoerythrin (PE, #562020), IL-17-PE (#559502), anti-human CD8-FITC (#555634), CD3-APC (#555342), IL-17-PE (#560436) (BD, USA); anti-CD3e (#BE0001-1), anti-CD28 (#BE0015-1), anti-IFN-γ (#BE0055), anti-IL-4 (#BE0045) (Bio X Cell, USA); IL-23 (#CS31), transforming growth factor-β1 (TGF-β1, #CA59), IL-6 (#CG39) (Novoprotein, China); anti-AHR (1:1000, #NB100-128, Novus, USA); anti-SGK1 (1:1000, #AF6789), anti-FOXO1 (1:1000, #AF6416), anti-p-FOXO1 (1:1000, #AF3417), anti-retinoic acid receptor-related orphan receptor gamma-t (RORγt; 1:1000, #DF3196), and anti-GAPDH (1:5000, #AF7021) (Affinity, USA).

Techniques: Staining, Flow Cytometry, Immunofluorescence

Journal: Military Medical Research

Article Title: High-salt-driven gut microbiota dysfunction aggravates prostatitis by promoting AHR/SGK1/FOXO1 axis-mediated Th17 cell differentiation

doi: 10.1186/s40779-025-00607-1

Figure Lengend Snippet: Inhibition of AHR abrogated the protective effects of 5-HIAA supplementation on EAP mice fed an HSD. a AHR levels in prostate tissues of mice among the Ctrl + NSD, Ctrl + HSD, EAP + NSD, EAP + HSD, and EAP + HSD + 5-HIAA groups ( n = 4). b A simple schematic diagram of the experimental workflow. c HE staining and inflammation scores of prostate tissues (scale bar = 100 μm, n = 4). d Tactile allodynia development in the mice from the Ctrl + NSD, Ctrl + HSD, EAP + NSD, EAP + HSD, EAP + HSD + 5-HIAA, and EAP + HSD + 5-HIAA + CH groups ( n = 4). e The levels of IL-1β, TNF-α, and IL-17A in the serum of the mice in the 6 groups ( n = 4). f Proportion of Th17 of CD4 + T cells, as determined by flow cytometry, among the splenic lymphocytes of immunized mice in the 6 groups (white arrowheads, n = 4). g The infiltration of Th17 cells in prostate tissues from mice was evaluated by immunofluorescence (scale bar = 100 μm, n = 4). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns non-significant. NOD nonobese male diabetic/LtJ, BF breeding feed, EAP experimental autoimmune prostatitis, HSD high-salt diet, NSD normal-salt diet, CH CH223191 (an AHR inhibitor), HE hematoxylin–eosin, IL-1β interleukin-1β, TNF-α tumor necrosis factor-α, IL-17A interleukin-17A, 5-HIAA 5-hydroxyindole acetic acid, AHR aryl hydrocarbon receptor, DAPI 4',6-diamidino-2-phenylindole

Article Snippet: The following reagents and antibodies were used: CFA (#F5881, Sigma, USA); commercial enzyme-linked immunosorbent assay (ELISA) kits for IFN-γ (#E-EL-M0048c), IL-1β (#E-EL-M0037), IL-17A (#E-EL-M0047c), tumor necrosis factor-α (TNF-α, #E-EL-M3063), and 5-HIAA (#E-EL-0075c) (Elabscience, China); anti-mouse CD4-fluorescein isothiocyanate (FITC, #553047), CD3e-allophycocyanin (APC, #561826), IFN-γ-phycoerythrin (PE, #562020), IL-17-PE (#559502), anti-human CD8-FITC (#555634), CD3-APC (#555342), IL-17-PE (#560436) (BD, USA); anti-CD3e (#BE0001-1), anti-CD28 (#BE0015-1), anti-IFN-γ (#BE0055), anti-IL-4 (#BE0045) (Bio X Cell, USA); IL-23 (#CS31), transforming growth factor-β1 (TGF-β1, #CA59), IL-6 (#CG39) (Novoprotein, China); anti-AHR (1:1000, #NB100-128, Novus, USA); anti-SGK1 (1:1000, #AF6789), anti-FOXO1 (1:1000, #AF6416), anti-p-FOXO1 (1:1000, #AF3417), anti-retinoic acid receptor-related orphan receptor gamma-t (RORγt; 1:1000, #DF3196), and anti-GAPDH (1:5000, #AF7021) (Affinity, USA).

Techniques: Inhibition, Staining, Flow Cytometry, Immunofluorescence

Journal: Military Medical Research

Article Title: High-salt-driven gut microbiota dysfunction aggravates prostatitis by promoting AHR/SGK1/FOXO1 axis-mediated Th17 cell differentiation

doi: 10.1186/s40779-025-00607-1

Figure Lengend Snippet: HSD promoted the differentiation of Th17 cells by activating SGK1/FOXO1 signaling pathway. a The in vitro Th17 cell differentiation ratio was determined by flow cytometry ( n = 3). b Th17 cell differentiation ratio was determined in an in vitro differentiation experiment using immunofluorescence (scale bar = 100 μm, n = 3). c SGK1, p-FOXO1, and FOXO1 levels in prostate tissues from Ctrl + NSD, Ctrl + HSD, EAP + NSD, EAP + HSD, and EAP + HSD + 5-HIAA mice ( n = 4). d AHR, SGK1, p-FOXO1, and FOXO1 levels in prostate tissues of mice in the EAP + HSD + 5-HIAA and EAP + HSD + 5-HIAA + CH groups ( n = 4). e AHR, SGK1, p-FOXO1, and FOXO1 levels in naïve CD4 + T cells of Ctrl, Sti, Sti + HSD, Sti + HSD + 5-HIAA, Sti + HSD + 5-HIAA + CH groups ( n = 3). f Transcriptional levels of Sgk1 in naïve CD4 + T cells of Ctrl, Sti, Sti + HSD, Sti + HSD + 5-HIAA, Sti + HSD + 5-HIAA + CH groups (n = 3). g For the ChIP-qPCR assay, 3 potential enhancer segments of the mouse promoter region of Sgk1 were created. h qPCR was used to assess the immunoprecipitated DNA by a ChIP assay ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns non-significant. EAP experimental autoimmune prostatitis, HSD high-salt diet, NSD normal-salt diet, IL-17A interleukin-17A, 5-HIAA 5-hydroxyindole acetic acid, AHR aryl hydrocarbon receptor, SGK1 serum and glucocorticoid-regulated kinase 1, FOXO1 forkhead box protein O1, CH CH223191 (an AHR inhibitor), ChIP chromatin immunoprecipitation, qPCR quantitative polymerase chain reaction, DAPI 4’,6-diamidino-2-phenylindole, Sti stimulation with IL-6, IL-23, TGF-β, anti-IFN-γ, and anti-IL-4

Article Snippet: The following reagents and antibodies were used: CFA (#F5881, Sigma, USA); commercial enzyme-linked immunosorbent assay (ELISA) kits for IFN-γ (#E-EL-M0048c), IL-1β (#E-EL-M0037), IL-17A (#E-EL-M0047c), tumor necrosis factor-α (TNF-α, #E-EL-M3063), and 5-HIAA (#E-EL-0075c) (Elabscience, China); anti-mouse CD4-fluorescein isothiocyanate (FITC, #553047), CD3e-allophycocyanin (APC, #561826), IFN-γ-phycoerythrin (PE, #562020), IL-17-PE (#559502), anti-human CD8-FITC (#555634), CD3-APC (#555342), IL-17-PE (#560436) (BD, USA); anti-CD3e (#BE0001-1), anti-CD28 (#BE0015-1), anti-IFN-γ (#BE0055), anti-IL-4 (#BE0045) (Bio X Cell, USA); IL-23 (#CS31), transforming growth factor-β1 (TGF-β1, #CA59), IL-6 (#CG39) (Novoprotein, China); anti-AHR (1:1000, #NB100-128, Novus, USA); anti-SGK1 (1:1000, #AF6789), anti-FOXO1 (1:1000, #AF6416), anti-p-FOXO1 (1:1000, #AF3417), anti-retinoic acid receptor-related orphan receptor gamma-t (RORγt; 1:1000, #DF3196), and anti-GAPDH (1:5000, #AF7021) (Affinity, USA).

Techniques: In Vitro, Cell Differentiation, Flow Cytometry, Immunofluorescence, ChIP-qPCR, Immunoprecipitation, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction

Journal: bioRxiv

Article Title: Au 18 (L-NIBC) 14 targets ASC oligomerization for autoimmune disease treatment

doi: 10.1101/2025.03.25.645153

Figure Lengend Snippet: a-c , ELISA results show that Au 18 (L-NIBC) 14 significantly reduced the levels of IL-1β ( a ), IL-18 ( b ), and caspase-1 ( c ) in the spinal cords of EAE mice, while Au 25 (L-NIBC) 18 showed no evidential inhibitory effects. d-e , Representative fluorescence microscopy images ( d ) and quantitative analysis ( e ) reveal a marked reduction in ASC specks in the spinal cords of EAE mice treated with Au 18 (L-NIBC) 14 , indicating effective inhibition of inflammasome activation. Au 25 (L-NIBC) 18 did not reduce the ratio of ASC specks. f-k . Regulation of other cytokines: pro-inflammatory cytokines TNF-α ( f ), IL- 17A ( g ) and IL-12 ( h ) were significantly reduced by Au 18 (L-NIBC) 14 ; while anti-inflammatory cytokines IL-4 ( i ), IL-10 ( j ) and IFN-γ ( k ) levels were significantly increased by Au 18 (L-NIBC) 14 .

Article Snippet: Methanol (HPLC, Thermo Fisher); NaOH (AR, Sinopharm), Ltd.; KH 2 PO 4 (AR, Sinopharm); NaH 2 PO 4 (AR, Sinopharm); Phorbol 12-myristate 13-acetate (HY-18739, MedChemExpress); Nigericin (HY-100381, MedChemExpress); poly(dA:dT) (tlrl-patn, InvivoGen); Flagellin (SRP8029, Sigma); MDP (M9508, Sigma); Lipofectamine TM 2000 Transfection Reagent (11668019, Thermo Fisher); Human IL-1β ELISA Kit (RK00001, ABclonal); Human IL-18 ELISA Kit(RK00176, ABclonal); Mouse IL-1β ELISA Kit (RK00006, ABclonal); Mouse IL-18 ELISA Kit (BMS618-3, Invitrogen); Mouse TNF-α ELISA Kit (RK00027, ABclonal); Mouse Caspase-1 ELISA Kit (AG-45B-0002-KI01, AdipoGen); Mouse IL-17A ELISA Kit (RK00039, ABclonal); Mouse IFN-γ ELISA Kit (RK00019, ABclonal); Mouse IL-4 ELISA Kit (RK00036, ABclonal); Mouse IL-10 ELISA Kit (RK00016, ABclonal); Mouse IL-12 Elisa Kit (E- EL-M3062, Elabscience); Hematoxylin and Eosin Staining Kit (C0105S, Beyotime); Luxol Fast Blue Stain Solution (G3242, Solarbio); BCA Protein Assay Kit (P0012, Beyotime); RIPA Lysis Buffer (P0013B, Beyotime); ECL (S6009L, US Everbright); Gota Serum (16210064, Thermo Fisher); Antifade Mounting Medium with DAPI (A4084, US Everbright); Rabbit anti-MBP antibody (78896T, Cell Signaling Technology); Rabbit anti-ASC antibody (67824, Cell Signaling Technology); Mouse anti-Occludin antibody (33-1500, Invitrogen); Mouse anti-GAPDH antibody (MA1- 16757, Invitrogen); Goat anti-mouse antibody (A0216, Beyotime); Alexa Fluor 568 (A- 11011, Thermo Fisher).

Techniques: Enzyme-linked Immunosorbent Assay, Fluorescence, Microscopy, Inhibition, Activation Assay

Journal: Mediators of Inflammation

Article Title: Notch Signaling Pathway Promotes Th17 Cell Differentiation and Participates in Thyroid Autoimmune Injury in Experimental Autoimmune Thyroiditis Mice

doi: 10.1155/2023/1195149

Figure Lengend Snippet: List of primers for qRT-PCR.

Article Snippet: SMC culture supernatants were collected, and the concentrations of IL-17A were measured using the mouse IL-17A ELISA kit (R&D Systems, USA) according to the manufacturer's instructions.

Techniques: Sequencing

Journal: Pharmaceutical biology

Article Title: Panax notoginseng saponins (PNS) attenuate Th17 cell differentiation in CIA mice via inhibition of nuclear PKM2-mediated STAT3 phosphorylation.

doi: 10.1080/13880209.2023.2173248

Figure Lengend Snippet: Figure 2. PNS reduced Th17 cell differentiation in vitro. (A,B) Representative dot plots and the percentage of Th17 cells (CD4þIL-17þ cells) (with the condition of Th17-polarization) treated with PNS (5, 10 and 20 lg/mL) were determined by flow cytometry. (C) The cell viability of CD4þT cells (with the condition of Th17-polar- ization) treated with PNS was detected by CCK8. (D) The mRNA expression of RORct in CD4þT cells (with the condition of Th17-polarization) treated with PNS (5, 10 and 20 lg/mL) was evaluated by RT-qPCR. (E) The levels of IL-17A in cell supernatants was determined by ELISA. Data were expressed as the mean ± SEM (n ¼ 3). p < 0.05, p < 0.01, p < 0.001 compared with the control group by one-way ANOVA with Tukey’s post hoc test.

Article Snippet: The levels of TGF-b, tumor necrosis factor-a (TNF-a, EK282HS), IL-6 (EK206HS), and interleukin 17A (IL-17A, EK217HS) in mouse serum and cell supernatants were measured by Mouse High Sensitivity ELISA kit (all for MultiSciences) according to the manufacturer’s instructions.

Techniques: Cell Differentiation, In Vitro, Flow Cytometry, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Control

Journal: Pharmaceutical biology

Article Title: Panax notoginseng saponins (PNS) attenuate Th17 cell differentiation in CIA mice via inhibition of nuclear PKM2-mediated STAT3 phosphorylation.

doi: 10.1080/13880209.2023.2173248

Figure Lengend Snippet: Figure 5. Th17 cell differentiation accompanied high PKM2 expression and nuclear accumulation. (A,B) Representative dot plots and the percentage of Th17 cells (CD4þIL-17þ cells) (with or without the condition of Th17-polarization) were determined by flow cytometry at different time points after culture. (C and E) The mRNA expression of RORct and PKM2 were evaluated by RT-qPCR in CD4þT cells (with or without the condition of Th17-polarization) at different time points after culture. (D) The levels of IL-17A in Th17 cells at different time points after culture were measured by ELISA. (F–H) CD4þT cells (with or without the condition of Th17-polariza- tion) were collected at different time points of differentiation, crosslinked with DSS, and analyzed for dimeric/tetrameric PKM2 expression by western blots. b-Actin was used as a loading control. (I,J) CD4þT cells (with or without the condition of Th17-polarization) were collected at different time points of differentiation. Nuclear and cytoplasmic fractions were isolated by cell fractionation and analyzed for PKM2 expression by western blots. Histone H3 and b-actin were used as nuclear and cytoplasm loading controls, respectively. (K,L) Subcellular localization of PKM2 were shown in left panel (scale bars, 10 lm). Cells were immunostained with anti-PKM2 (PKM2, red). The nucleus was marked with DAPI (blue). Merged images (Merge) were shown. The framed regions were zoomed in the fourth row (Zoom). The scatter diagram of PKM2 and DAPI signals were measured by Fiji ImageJ software. The co-localization of PKM2 and nucleus was expressed as person correlation coefficient determined by Fiji ImageJ software. Data were expressed as the mean ± SEM (n ¼ 3). p < 0.05, p < 0.01, p < 0.001 compared with the control group by one- way ANOVA with Tukey’s post hoc test.

Article Snippet: The levels of TGF-b, tumor necrosis factor-a (TNF-a, EK282HS), IL-6 (EK206HS), and interleukin 17A (IL-17A, EK217HS) in mouse serum and cell supernatants were measured by Mouse High Sensitivity ELISA kit (all for MultiSciences) according to the manufacturer’s instructions.

Techniques: Cell Differentiation, Expressing, Flow Cytometry, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Western Blot, Control, Isolation, Cell Fractionation, Software

Journal: Pharmaceutical biology

Article Title: Panax notoginseng saponins (PNS) attenuate Th17 cell differentiation in CIA mice via inhibition of nuclear PKM2-mediated STAT3 phosphorylation.

doi: 10.1080/13880209.2023.2173248

Figure Lengend Snippet: Figure 6. PKM2 nuclear accumulation promoted Th17 cell differentiation in vitro. (A,B) Representative dot plots and the percentage of Th17 cells (CD4þIL-17þ cells) (with the condition of Th17-polarization) treated with Tepp-46 (50, 100 and 150 lM) were determined by flow cytometry. (C) The mRNA expression of RORct in CD4þT cells (with the condition of Th17-polarization) treated with Tepp-46 (50, 100 and 150lM) was evaluated by RT-qPCR. (D) The IL-17A levels in medium of CD4þT cells (with the condition of Th17-polarization) treated with Tepp-46 (50, 100 and 150lM) was detected by ELISA. (E,F) Representative dot plots and the per- centage of Th17 cells (CD4þIL-17þ cells) (with the condition of Th17-polarization) treated with SAICAR (2, 4 and 8 lM) were determined by flow cytometry. (G) The mRNA expression of RORct in CD4þT cells (with the condition of Th17-polarization) treated with SAICAR (2, 4 and 8 lM) was evaluated by RT-qPCR. (H) The IL-17A levels in medium of CD4þT cells (with the condition of Th17-polarization) treated with SAICAR (2, 4 and 8 lM) was detected by ELISA. Data were expressed as the mean ± SEM (n ¼ 3). p < 0.05, p < 0.01, p < 0.001 compared with the control group by one-way ANOVA with Tukey’s post hoc test.

Article Snippet: The levels of TGF-b, tumor necrosis factor-a (TNF-a, EK282HS), IL-6 (EK206HS), and interleukin 17A (IL-17A, EK217HS) in mouse serum and cell supernatants were measured by Mouse High Sensitivity ELISA kit (all for MultiSciences) according to the manufacturer’s instructions.

Techniques: Cell Differentiation, In Vitro, Flow Cytometry, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Control