Journal: Research

Article Title: Pim1 Serves as a Therapeutic Target for Inflammatory Arthritis via Mitochondrial Metabolism and Th17 Cell Differentiation

doi: 10.34133/research.1137

Figure Lengend Snippet: Specifically intercepting Pim1 alleviates the development of inflammatory arthritis by impeding Th17 cell differentiation. (A) Identification of Pim1 flox and CD4-cre gene. WT, wild type. (B) Relative protein levels of Pim1 in CD4 + T cells isolated from Pim1 flox and Pim1 cKO mice. (C) Macroscopic images of the ankles, arthritis scores, and hind paw thicknesses of Pim1 flox and Pim1 cKO CIA mice ( n = 5). (D and E) Representative histological images with H&E staining (D) and safranin O-fast green staining (E) of ankles from Pim1 flox and Pim1 cKO CIA mice ( n = 10). Scale bars, 200 μm. (F) Representative micro-CT images of the ankles of Pim1 flox and Pim1 cKO CIA mice. (G) Frequencies of IFN-γ + , IL-4 + , IL-17A + , and Foxp3 + cells among CD4 + cells in the spleens of Pim1 flox and Pim1 cKO CIA mice ( n = 5). (H) Relative mRNA expression of RORγt, T-bet, GATA3, and Foxp3 in the ankles of Pim1 flox or Pim1 cKO CIA mice ( n = 5). (I) Frequencies of IL-17A + cells among CD4 + cells in the ankles of Pim1 flox and Pim1 cKO CIA mice ( n = 5). (J) Relative mRNA expression of IL-17A in the ankles of Pim1 flox or Pim1 cKO CIA mice ( n = 5). (K) Representative immunohistochemical (IHC) images showing IL-17A expression in the ankle tissues of Pim1 flox and Pim1 cKO CIA mice. Scale bars, 50 μm. The data were statistically analyzed via 2-tailed Student’s t test (B, D, E, and G to J) and 2-way repeated-measures ANOVA (C).

Article Snippet: Human IL-17A enzyme-linked immunosorbent assay (ELISA) kit (Elabscience, #E-EL-H5812), human IL-17F ELISA kit (Elabscience, #E-EL-H4193), human IL-22 ELISA kit (Elabscience, #E-EL-H0106), and human GM-CSF ELISA kit (Elabscience, #E-EL-H0081) were used.

Techniques: Cell Differentiation, Isolation, Staining, Micro-CT, Expressing, Immunohistochemical staining

Journal: Research

Article Title: Pim1 Serves as a Therapeutic Target for Inflammatory Arthritis via Mitochondrial Metabolism and Th17 Cell Differentiation

doi: 10.34133/research.1137

Figure Lengend Snippet: Pim1 promotes Th17 cell differentiation in vitro. (A) Relative mRNA levels of Pim1 during Th17 cell differentiation ( n = 9). (B) Relative protein levels of pSTAT3, RORγt, and Pim1 during Th17 cell differentiation ( n = 9). (C and D) Frequency of Th17 cells among CD4 + cells after AZD1208 treatment ( n = 9). (E) Relative protein levels of Pim1 among CD4 + T cells overexpressing (OE) vector or Pim1 ( n = 9). (F) Frequency of Th17 cells in CD4 + cells after overexpressing vector or Pim1 ( n = 9). (G) Relative protein levels of RORγt and pSTAT3 after AZD1208 treatment or Pim1 overexpression ( n = 9). (H and I) Relative mRNA levels of Th17-cell-associated pathogenic genes after AZD1208 treatment (H) or Pim1 overexpression (I) ( n = 9). (J and K) Concentration of IL-17A, IL-17F, IL-22, and GM-CSF in the cell supernatant after AZD1208 treatment (J) or Pim1 overexpression (K). (L) Frequencies of Th1, Th2, and Treg cells among CD4 + T cells after treatment with AZD1208 ( n = 9). The data were statistically analyzed via one-way ANOVA, followed by Bonferroni’s post hoc comparisons (A to D) and paired t test (E to L).

Article Snippet: Human IL-17A enzyme-linked immunosorbent assay (ELISA) kit (Elabscience, #E-EL-H5812), human IL-17F ELISA kit (Elabscience, #E-EL-H4193), human IL-22 ELISA kit (Elabscience, #E-EL-H0106), and human GM-CSF ELISA kit (Elabscience, #E-EL-H0081) were used.

Techniques: Cell Differentiation, In Vitro, Plasmid Preparation, Over Expression, Concentration Assay

Journal: Research

Article Title: Pim1 Serves as a Therapeutic Target for Inflammatory Arthritis via Mitochondrial Metabolism and Th17 Cell Differentiation

doi: 10.34133/research.1137

Figure Lengend Snippet: Nilotinib inhibits Th17 cell differentiation and alleviates inflammatory arthritis by targeting Pim1. (A) Enzyme activity of Pim1 after incubation with AZD1208, drospirenone, olaparib, nilotinib, or dutasteride ( n = 9). (B) Frequency of Th17 cells among CD4 + cells after treatment with nilotinib in the presence or absence of Pim1 overexpression ( n = 9). (C) MFI of Rhod-2AM in cells treated with nilotinib in the presence or absence of Pim1 overexpression ( n = 9). (D) OCR of cells treated with nilotinib in the presence or absence of Pim1 overexpression ( n = 9). (E) Macroscopic images of the ankles of Pim1 flox and Pim1 cKO CIA mice treated with vehicle or nilotinib. (F) Arthritis scores and hind paw thickness of Pim1 flox and Pim1 cKO CIA mice treated with vehicle or nilotinib ( n = 5). (G and H) Representative histological images with H&E staining (G) and safranin O-fast green staining (H) of the ankles of Pim1 flox and Pim1 cKO CIA mice treated with vehicle or nilotinib ( n = 10). Scale bars, 200 μm. (I) Representative micro-CT images of ankles of Pim1 flox and Pim1 cKO CIA mice treated with vehicle or nilotinib. (J and K) Frequency of Th17 cells among CD4 + cells in spleens (J) and ankles (K) of Pim1 flox and Pim1 cKO CIA mice treated with vehicle or nilotinib ( n = 5). (L) Representative immunohistochemical images showing IL-17A expression in the ankle tissues of Pim1 flox and Pim1 cKO CIA mice treated with vehicle or nilotinib. Scale bars, 50 μm. The data were statistically analyzed via one-way ANOVA, followed by Bonferroni’s post hoc comparisons (B to D, G, H, J, and K) and 2-way repeated-measures ANOVA (F).

Article Snippet: Human IL-17A enzyme-linked immunosorbent assay (ELISA) kit (Elabscience, #E-EL-H5812), human IL-17F ELISA kit (Elabscience, #E-EL-H4193), human IL-22 ELISA kit (Elabscience, #E-EL-H0106), and human GM-CSF ELISA kit (Elabscience, #E-EL-H0081) were used.

Techniques: Cell Differentiation, Activity Assay, Incubation, Over Expression, Staining, Micro-CT, Immunohistochemical staining, Expressing

Journal: Research

Article Title: Pim1 Serves as a Therapeutic Target for Inflammatory Arthritis via Mitochondrial Metabolism and Th17 Cell Differentiation

doi: 10.34133/research.1137

Figure Lengend Snippet: Nilotinib acts as an effective drug for the treatment of inflammatory arthritis. (A) Macroscopic images of the ankles, arthritis scores, and hind paw thickness of CIA mice treated with vehicle (NC group) and nilotinib (nilotinib group) ( n = 5). (B) Macroscopic images of the ankles, arthritis scores, and incidence rate of SKG arthritis in the NC and nilotinib groups ( n = 5). (C and D) Representative histological images with H&E staining (C) and safranin O-fast green staining (D) of the ankles of CIA and SKG arthritis mice in the NC and nilotinib groups ( n = 10). Scale bars, 200 μm. (E) Representative micro-CT images of the ankles of CIA and SKG arthritis mice in the NC and nilotinib groups. (F and G) Frequency of Th17 cells among CD4 + cells in the spleens (F) and ankles (G) of CIA and SKG arthritis mice in the NC and nilotinib groups ( n = 5). (H) Representative immunohistochemical images showing IL-17A expression in the ankle tissues of CIA and SKG arthritis mice in the NC and nilotinib groups. Scale bars, 50 μm. The data were statistically analyzed via 2-way repeated-measures ANOVA (A and B) and 2-tailed Student’s t test (C, D, F, and G).

Article Snippet: Human IL-17A enzyme-linked immunosorbent assay (ELISA) kit (Elabscience, #E-EL-H5812), human IL-17F ELISA kit (Elabscience, #E-EL-H4193), human IL-22 ELISA kit (Elabscience, #E-EL-H0106), and human GM-CSF ELISA kit (Elabscience, #E-EL-H0081) were used.

Techniques: Staining, Micro-CT, Immunohistochemical staining, Expressing