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: PLOS One

Article Title: NFIC suppressed the development of Glioma via modulating the balance of SHP2/PI3K and NF-κB/PTEN Signaling

doi: 10.1371/journal.pone.0341816

Figure Lengend Snippet: NFIC promotes OGN and PTEN expression while inhibiting NF-κB, SHP2, and phosphorylated SHP2 expression. A: Western blot analysis of NFIC, OGN, p-NF-κB, SHP2, and p-SHP2 protein bands in six groups (NFIC-NC, NFIC-OE, NF-κB NC, NF-κB NC, PHPS1 NC, PHPS1 OE), with statistical analysis of relative protein expression levels. B: Western blot analysis of PTEN and HEY1 protein bands in six groups, along with statistical analysis of relative protein expression levels. Data are expressed as mean ± standard deviation. N = 3, P < 0.05 indicates statistically significant difference, * P < 0.05; ** P < 0.01; nsP > 0.05.

Article Snippet: The membranes were blocked in TBST buffer containing 5% skimmed milk at 37°C for 2 hours, followed by overnight incubation at 4°C with the following primary antibodies: NFIC (CST, #11911, 1:1000), OGN (CST, #24083, 1:1000), NF-κB (CST, #8242, 1:1000), SHP2 (CST, #3752, 1:1000), p-SHP2 (CST, #13379, 1:1000), PI3K (CST, #4292, 1:1000), AKT (CST, #4060, 1:2000), Cyclin A1 (CST, #4656, 1:2000), Cyclin D1 (CST, #2922, 1:1000), MMP-3 (CST, #14351, 1:1000), and MMP-9 (CST, #3852, 1:1000).

Techniques: Expressing, Western Blot, Standard Deviation

Journal: PLOS One

Article Title: NFIC suppressed the development of Glioma via modulating the balance of SHP2/PI3K and NF-κB/PTEN Signaling

doi: 10.1371/journal.pone.0341816

Figure Lengend Snippet: NFIC promotes OGN and PTEN expression while suppressing NF-κB, SHP2, and phosphorylated SHP2 expression. A: Western blot detection of p-PIK, p-AKT, p-STAT3, GAPDH protein bands, and statistical analysis of relative protein expression levels; B: Western blot detection of Cyclin A1, Cyclin D1, MMP-3, and MMP-9 protein bands in the six groups, and statistical analysis of relative protein expression levels. Data are expressed as mean ± standard deviation. N = 3, P < 0.05 indicates statistically significant difference, * P < 0.05; ** P < 0.01; nsP > 0.05.

Article Snippet: The membranes were blocked in TBST buffer containing 5% skimmed milk at 37°C for 2 hours, followed by overnight incubation at 4°C with the following primary antibodies: NFIC (CST, #11911, 1:1000), OGN (CST, #24083, 1:1000), NF-κB (CST, #8242, 1:1000), SHP2 (CST, #3752, 1:1000), p-SHP2 (CST, #13379, 1:1000), PI3K (CST, #4292, 1:1000), AKT (CST, #4060, 1:2000), Cyclin A1 (CST, #4656, 1:2000), Cyclin D1 (CST, #2922, 1:1000), MMP-3 (CST, #14351, 1:1000), and MMP-9 (CST, #3852, 1:1000).

Techniques: Expressing, Western Blot, Standard Deviation

Journal: PLOS One

Article Title: NFIC suppressed the development of Glioma via modulating the balance of SHP2/PI3K and NF-κB/PTEN Signaling

doi: 10.1371/journal.pone.0341816

Figure Lengend Snippet: NFIC inhibits glioblastoma cell proliferation and invasion, while NF-κB promotes these processes. A: Co-immunoprecipitation (CO-IP) analysis of NFIC and PTEN protein bands in six sample groups; B: CO-IP analysis of OGN and NF-κB protein bands in six sample groups; C: CO-IP analysis of NF-κB and SHP2 protein bands in six sample groups; D: CO-IP analysis of NFIC and OGN protein bands in six sample groups; Data are expressed as mean ± standard deviation. N = 3, P < 0.05 indicates statistically significant difference; * P < 0.05; ** P < 0.01; nsP > 0.05.

Article Snippet: The membranes were blocked in TBST buffer containing 5% skimmed milk at 37°C for 2 hours, followed by overnight incubation at 4°C with the following primary antibodies: NFIC (CST, #11911, 1:1000), OGN (CST, #24083, 1:1000), NF-κB (CST, #8242, 1:1000), SHP2 (CST, #3752, 1:1000), p-SHP2 (CST, #13379, 1:1000), PI3K (CST, #4292, 1:1000), AKT (CST, #4060, 1:2000), Cyclin A1 (CST, #4656, 1:2000), Cyclin D1 (CST, #2922, 1:1000), MMP-3 (CST, #14351, 1:1000), and MMP-9 (CST, #3852, 1:1000).

Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Standard Deviation

Journal: PLOS One

Article Title: NFIC suppressed the development of Glioma via modulating the balance of SHP2/PI3K and NF-κB/PTEN Signaling

doi: 10.1371/journal.pone.0341816

Figure Lengend Snippet: NFIC binds to the promoter regions of OGN and PTEN and regulates their transcription, leading to increased expression of these two genes. Immunohistochemical staining results for NFIC, PTEN, OGN, NF-κB, and p-SHP2, along with statistical analysis of staining. Data are presented as mean ± standard deviation. N = 3, P < 0.05 indicates statistically significant difference, *P < 0.05, **P < 0.01.

Article Snippet: The membranes were blocked in TBST buffer containing 5% skimmed milk at 37°C for 2 hours, followed by overnight incubation at 4°C with the following primary antibodies: NFIC (CST, #11911, 1:1000), OGN (CST, #24083, 1:1000), NF-κB (CST, #8242, 1:1000), SHP2 (CST, #3752, 1:1000), p-SHP2 (CST, #13379, 1:1000), PI3K (CST, #4292, 1:1000), AKT (CST, #4060, 1:2000), Cyclin A1 (CST, #4656, 1:2000), Cyclin D1 (CST, #2922, 1:1000), MMP-3 (CST, #14351, 1:1000), and MMP-9 (CST, #3852, 1:1000).

Techniques: Expressing, Immunohistochemical staining, Staining, Standard Deviation

Journal: PLOS One

Article Title: NFIC suppressed the development of Glioma via modulating the balance of SHP2/PI3K and NF-κB/PTEN Signaling

doi: 10.1371/journal.pone.0341816

Figure Lengend Snippet: NFIC promotes OGN and PTEN expression while suppressing NF-κB, SHP2, p-SHP2, PI3K, AKT, Cyclin A1, Cyclin D1, MMP-3, and MMP-9 expression. NF-κB promotes SHP2 expression, while OGN and PTEN inhibit p-SHP2 expression. NFIC suppresses glioma cell proliferation and invasion, whereas NF-κB promotes these processes.

Article Snippet: The membranes were blocked in TBST buffer containing 5% skimmed milk at 37°C for 2 hours, followed by overnight incubation at 4°C with the following primary antibodies: NFIC (CST, #11911, 1:1000), OGN (CST, #24083, 1:1000), NF-κB (CST, #8242, 1:1000), SHP2 (CST, #3752, 1:1000), p-SHP2 (CST, #13379, 1:1000), PI3K (CST, #4292, 1:1000), AKT (CST, #4060, 1:2000), Cyclin A1 (CST, #4656, 1:2000), Cyclin D1 (CST, #2922, 1:1000), MMP-3 (CST, #14351, 1:1000), and MMP-9 (CST, #3852, 1:1000).

Techniques: Expressing