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: Cell Reports

Article Title: De novo steroidogenesis in tumor cells drives bone metastasis and osteoclastogenesis

doi: 10.1016/j.celrep.2024.113936

Figure Lengend Snippet: Pregnenolone promotes human osteoclastogenesis and is secreted by human tumor cells (A–C) Representative images (A) and quantification (B and C) of TRAP-stained cell cultures of human blood mononuclear cell-derived OCs cultured for 2 days in the presence of 20 ng/mL recombinant human M-CSF and 20 ng/mL soluble human RANKL and then in the presence of the indicated concentration of pregnenolone or vehicle for 12 days. Scale bars represent 100 μm. (D) Levels of pregnenolone in the supernatants of A375, MDA-MB-435S, A549, NCI-H1299, NCI-H460, MCF-7, MDA-MB-231, DU145, LNCaP, PC-3, Hs895.T, SK-MEL-28, BEAS-2B, NCI-H128, NCI-H2126, HCC70, MCF-10A, MDA-PCa-2b, RWPE-1, and LASCPC-01 cells measured by ELISA. (E) Kaplan-Meier curve of breast-cancer-specific survival for 2,976 patients with high or low expression of Cyp11a1 in the primary tumor. Bar graphs show mean and SD of data from 3 independent experiments. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.002; n.s., not significant.

Article Snippet: Mouse Competitive ELISA Kit M-CSF , Elabscience Biotech. , Cat# E-EL-M2445.

Techniques: Staining, Derivative Assay, Cell Culture, Recombinant, Concentration Assay, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Cell Reports

Article Title: De novo steroidogenesis in tumor cells drives bone metastasis and osteoclastogenesis

doi: 10.1016/j.celrep.2024.113936

Figure Lengend Snippet:

Article Snippet: Mouse Competitive ELISA Kit M-CSF , Elabscience Biotech. , Cat# E-EL-M2445.

Techniques: Virus, shRNA, Recombinant, Modification, Staining, RNA Library Preparation, TaqMan Assay, Competitive ELISA, RNA Sequencing, Plasmid Preparation, Software, Real-time Polymerase Chain Reaction

Journal: Acta biomaterialia

Article Title: Spatial targeting of fibrosis-promoting macrophages with nanoscale metal-organic frameworks for idiopathic pulmonary fibrosis therapy.

doi: 10.1016/j.actbio.2023.12.006

Figure Lengend Snippet: Fig. 1. Overexpression of CSF-1R in CD206+ M2 macrophages. (A) Flow cytometry analysis of CSF-1R and CD206 expression in F4/80+ macrophages. Right panels: Quantified data in sorted F4/80+ macrophages. Results are expressed as means ± SD ( n = 3; ∗∗P < 0.01). (B) Representative results for coimmunostaining of CD206 and CSF-1R in the lung sections from bleomycin-treated mice. Representative images are shown. Bar = 20 μm. (C) Immunohistochemistry staining of CSF-1R and CD206 in the lung sections of IPF patients. Representative images are shown. Bar = 50 μm. (D-F) Single cell atlas of IPF patients according to dataset GSE122960. (D) Upper panels: Cellular populations identified. Lower panels: t-distributed stochastic neighbor embedding (t-SNE) depicting cell clusters originating either from a donor or from IPF patients. (E) Expression of CSF-1R for the cell types defined above each panel. (F) Percentage of cells with non-zero CSF-1R expression. (G) Kaplan– Meier survival analyses of IPF patients based on the expression of CSF-1R according to dataset GSE70866. (H) Immunofluorescence staining of CSF-1R and α-SMA on mouse lung tissues. Representative images are shown. Bar = 50 μm.

Article Snippet: The primary antibodies used were rabbit nti-phospho-CSF-1R, mouse anti- β-actin, and rabbit anti-GAPDH. horseradish peroxidase-conjugated goat anti-mouse/rabbit IgG Boster no. BA1050/BA1056, Wuhan, China, 1:10,0 0 0 dilution) was sed as the secondary antibody.

Techniques: Over Expression, Flow Cytometry, Expressing, Immunohistochemistry, Staining

Journal: Acta biomaterialia

Article Title: Spatial targeting of fibrosis-promoting macrophages with nanoscale metal-organic frameworks for idiopathic pulmonary fibrosis therapy.

doi: 10.1016/j.actbio.2023.12.006

Figure Lengend Snippet: Fig. 5. (A) Schematic diagram of the BLZ-loaded NPs therapy procedure. (B) Inhibitory effects of scrNP-BLZ@Mn-Cur or M2NP-BLZ@Mn-Cur on CSF-1R phosphorylation in lung tissues. (C) Representative flow cytometry results is shown to identify pulmonary macrophage subsets from lung digests in BLM-treated mice. Right panels: Quantified data in sorted F4/80+ macrophages ( n = 3, means ± SD). (D) Cell differential from BAL fluid was counted by Diff-Quik staining ( n = 5; mean ± SD; ∗p < 0.05). (E) The level of TGF- β1, IL-6, IL-11, and TNF- α in lung tissues were determined by ELISA. Results are expressed as means ± SD ( n = 3; ∗∗p < 0.01, ∗p < 0.05).

Article Snippet: The primary antibodies used were rabbit nti-phospho-CSF-1R, mouse anti- β-actin, and rabbit anti-GAPDH. horseradish peroxidase-conjugated goat anti-mouse/rabbit IgG Boster no. BA1050/BA1056, Wuhan, China, 1:10,0 0 0 dilution) was sed as the secondary antibody.

Techniques: Phospho-proteomics, Cytometry, Diff-Quik, Staining, Enzyme-linked Immunosorbent Assay