Journal: Redox Biology

Article Title: Hyodeoxycholic acid attenuates atherosclerosis by antagonizing FXR and modulating the PD-1/mTORC1 signaling axis

doi: 10.1016/j.redox.2026.104096

Figure Lengend Snippet: HDCA modulates Treg migration and atherosclerotic plaque composition via FXR signaling. ApoE−/− mice (C57BL/6J background, male, 8 weeks old) were fed a high-fat diet for 28 days to induce AS and subsequently received adoptive transfer of control or FXR-knockout (FXR KO) Treg cells generated by CRISPR/Cas9-mediated lentiviral transduction, with HDCA (30 μM) or vehicle treatment as indicated. (A) Representative Western blot analysis of FXR, PD-1, SHP-2, p-Raptor, RAC and IL-10R expression in isolated Treg cells from each group. (B) Oil Red O staining was performed to assess lipid accumulation in the aorta. (C) Masson's trichrome staining of aortic sections from FXR KO mice reveals comparable plaque area and collagen deposition in both HDCA-treated and untreated groups (magnification, 5 × ; scale bar, 1 mm). (D) Representative H&E images of aortic sections show the difference between untreated and HDCA-treated mice in the FXR KO groups (magnification, 5 × ; scale bar, 500 μm). Relative bar graphs show quantification of lesion area and lesion/media area ratio. (E) Confocal immunofluorescence was used to evaluate Foxp3+ Treg infiltration within atherosclerotic plaques (scale bar, 25 μm). (F) Immunohistochemistry images show Treg accumulation in the plaque area across all groups (magnification, 40 × ; scale bar, 100 μm). (G) Flow cytometry analysis of Treg proportions in aortic plaques and spleen from control and FXR KO mice, with or without HDCA treatment. (H) Western blot analysis of matrix remodeling-related proteins, including calpain 1 and matrix metalloproteinase 2, and the anti-inflammatory factor IL-10. Data are presented as mean ± SD (n = 5 biological replicates). Data with four groups were analyzed by one-way ANOVA with Tukey's post hoc test. Comparisons between two groups were performed using the non-parametric Mann-Whitney U test. ns, not significant; ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Proteins were detected using the following antibodies: anti-CPT1a antibody (ab234111, abcam), anti-beta actin antibody (ab8226, abcam), anti-PERK antibody (ab229912, abcam), anti-ERK1+ERK2 antibody (ab184699, abcam), anti-S6K1 antibody (ab14708, abcam), anti-S6K1 (phospho T229) antibody (ab5231, abcam), Rac1/2/3 antibody (G-2) (sc-514583, Santa Cruz), anti-Calpain 1 antibody (ab108400, abcam), anti-MMP2 antibody (ab92536, abcam), anti-IL-10 antibody (ab310329, abcam), anti-ZNF671 antibody (HPA046099, Sigma-Aldrich), anti-MAPK6/ERK3 antibody (ab53277, abcam), SIAH1 recombinant rabbit monoclonal antibody (PSH01-80) (MA5-51926, Thermo Fisher), p-Stat1 antibody (pY701.4A) (sc-136229, Santa Cruz), Stat1 antibody (C-136) (sc-464, Santa Cruz), anti-FXR1 antibody (ab155124, abcam), phospho-Raptor (Ser792) polyclonal antibody (PA5-118730, Thermo Fisher), anti-PD1 antibody (ab214421, abcam), SHP-2 antibody (3752S, Cell Signaling Technology), IL-10R antibody (3F9) (sc-53654, Santa Cruz), GAPDH antibody (6C5) (sc-32233, Santa Cruz), rabbit anti-mouse IgG H&L (HRP) (ab6728, abcam), goat anti-rabbit IgG (H + L) highly cross-adsorbed secondary antibody, Alexa FluorTM Plus 488 (A32731, Thermo Fisher).

Techniques: Migration, Adoptive Transfer Assay, Control, Knock-Out, Generated, CRISPR, Transduction, Western Blot, Expressing, Isolation, Staining, Immunofluorescence, Immunohistochemistry, Flow Cytometry, MANN-WHITNEY

Journal: Cancers

Article Title: Evidence for a Tumor-Suppressive Role of SHP-1 in EMT Regulation in Bladder Cancer Cells

doi: 10.3390/cancers18091401

Figure Lengend Snippet: ( A ) Relative SHP-1 protein expression in the 19 BCa cell lines examined in this study. ( B ) Expression levels were significantly higher in epithelial-like cells (n = 11) than in intermediate and mesenchymal-like lines (n = 8). Error bars represent the standard error of the mean (SEM).

Article Snippet: Lentiviral shRNA constructs targeting SHP-1 were obtained from Origene (Rockville, MD, USA), and the manufacturer’s protocol was used to establish stable cell lines for each construct.

Techniques: Expressing

Journal: Cancers

Article Title: Evidence for a Tumor-Suppressive Role of SHP-1 in EMT Regulation in Bladder Cancer Cells

doi: 10.3390/cancers18091401

Figure Lengend Snippet: ( A ) SHP-1 protein expression in bladder tissue was significantly lower in muscle-invasive (MI, n = 8) tumor than in non-muscle-invasive (NMI, n = 8) and urothelial tissue with no residual disease (NRD, n = 10). ( B ) Examples of DAB and corresponding H&E staining in the bladder tissues examined in this study. The error bars represent the interquartile range.

Article Snippet: Lentiviral shRNA constructs targeting SHP-1 were obtained from Origene (Rockville, MD, USA), and the manufacturer’s protocol was used to establish stable cell lines for each construct.

Techniques: Expressing, Staining

Journal: Cancers

Article Title: Evidence for a Tumor-Suppressive Role of SHP-1 in EMT Regulation in Bladder Cancer Cells

doi: 10.3390/cancers18091401

Figure Lengend Snippet: Correlation plots (n = 19) of SHP-1 protein expression with the EMT markers E-cadherin, N-cadherin, and Vimentin. Gray areas represent the 95%CI. Representative Western blot images: from the BCa cell lines EJ and HT-1376, the first lane of each pair corresponds to the protein indicated above each lane, and the second corresponds to the total protein detected for the same lane.

Article Snippet: Lentiviral shRNA constructs targeting SHP-1 were obtained from Origene (Rockville, MD, USA), and the manufacturer’s protocol was used to establish stable cell lines for each construct.

Techniques: Expressing, Western Blot

Journal: Cancers

Article Title: Evidence for a Tumor-Suppressive Role of SHP-1 in EMT Regulation in Bladder Cancer Cells

doi: 10.3390/cancers18091401

Figure Lengend Snippet: ( A ) SHP-1 protein expression following transduction with SHP-1 shRNA (shSHP-1) or SHP-1 cDNA (+SHP-1) and corresponding negative controls (NC): CUBIII n = 6, RT-112 n = 7, TCCSUP n = 4, UM-UC-3 n = 6. ( B ) Representative Western blot images: the first lane of each pair corresponds to SHP-1, and the second corresponds to the total protein detected for the same lane. Error bars represent the SEM. ( C ) Relative proliferation rates of BCa cells transduced with SHP-1 shRNA (shSHP-1) or SHP-1 cDNA (+SHP-1) and corresponding negative controls (NC): CUBIII n = 5, RT-112 n = 5, TCCSUP n = 9, UM-UC-3 n = 5. Error bars represent the SEM.

Article Snippet: Lentiviral shRNA constructs targeting SHP-1 were obtained from Origene (Rockville, MD, USA), and the manufacturer’s protocol was used to establish stable cell lines for each construct.

Techniques: Expressing, Transduction, shRNA, Western Blot

Journal: Cancers

Article Title: Evidence for a Tumor-Suppressive Role of SHP-1 in EMT Regulation in Bladder Cancer Cells

doi: 10.3390/cancers18091401

Figure Lengend Snippet: ( A ) Relative migration rates of BCa cells transduced with SHP-1 shRNA (shSHP-1) or SHP-1 cDNA (+SHP-1) and corresponding negative controls (NC): CUBIII n = 5, RT-112 n = 4, TCCSUP n = 3, UM-UC-3 n = 3. ( B ) Representative images from in vitro migration assays (scale bar = 200 μm). ( C ) Relative invasion rates of BCa cells transduced with SHP-1 shRNA (shSHP-1) or SHP-1 cDNA (+SHP-1) and corresponding negative controls (NC): CUBIII n = 3, RT-112 n = 5, TCCSUP n = 3, UM-UC-3 n = 3. ( D ) Representative images from in vitro invasion assays (scale bar = 200 μm). Error bars represent the SEM.

Article Snippet: Lentiviral shRNA constructs targeting SHP-1 were obtained from Origene (Rockville, MD, USA), and the manufacturer’s protocol was used to establish stable cell lines for each construct.

Techniques: Migration, Transduction, shRNA, In Vitro

Journal: Cancers

Article Title: Evidence for a Tumor-Suppressive Role of SHP-1 in EMT Regulation in Bladder Cancer Cells

doi: 10.3390/cancers18091401

Figure Lengend Snippet: Significant gene sets resulting from GSEA of RNA sequence data comparing the transduced BCa lines with high-SHP-1-expressing lines versus low-SHP-1-expressing lines (q-value < 0.1).

Article Snippet: Lentiviral shRNA constructs targeting SHP-1 were obtained from Origene (Rockville, MD, USA), and the manufacturer’s protocol was used to establish stable cell lines for each construct.

Techniques: Sequencing, Expressing

Journal: Cancers

Article Title: Evidence for a Tumor-Suppressive Role of SHP-1 in EMT Regulation in Bladder Cancer Cells

doi: 10.3390/cancers18091401

Figure Lengend Snippet: Relative expression of pAkt/Akt in BCa cells transduced with SHP-1 shRNA (shSHP-1) or SHP-1 cDNA (+SHP-1) and corresponding negative controls (NC): RT-112 n = 7 and TCCSUP n = 8. Representative Western blot images: the first lane of each pair corresponds to Akt or pAkt (ser473), and the second corresponds to the total protein detected for the same lane. Error bars represent the SEM.

Article Snippet: Lentiviral shRNA constructs targeting SHP-1 were obtained from Origene (Rockville, MD, USA), and the manufacturer’s protocol was used to establish stable cell lines for each construct.

Techniques: Expressing, Transduction, shRNA, Western Blot