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FXR and HDCA regulates the levels of intestinal inflammation. (A) The relative expression of mRNA level in the distal ileum (n = 6). (B) Representative immunoblots of proteins in the distal ileum (n = 3). Relative expression levels of NLRP3 (C), IL-6 <t>(D),</t> <t>TNF-α</t> (E), IL-18 (F), IL-10 (G), IL-1β p17 (H). (I) Immunohistochemical staining and (J) quantitative analysis of NLRP3+, <t>IL-6+,</t> <t>TNF-α+</t> and IL-18+ cells in the distal ileum (scale bar, 100 μm, n = 6). * P < 0.05, ** P < 0.01 by One-way ANOVA with post hoc Tukey's test (A, C–H, J). Data is presented as mean ± SEM.
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FXR and HDCA regulates the levels of intestinal inflammation. (A) The relative expression of mRNA level in the distal ileum (n = 6). (B) Representative immunoblots of proteins in the distal ileum (n = 3). Relative expression levels of NLRP3 (C), IL-6 <t>(D),</t> <t>TNF-α</t> (E), IL-18 (F), IL-10 (G), IL-1β p17 (H). (I) Immunohistochemical staining and (J) quantitative analysis of NLRP3+, <t>IL-6+,</t> <t>TNF-α+</t> and IL-18+ cells in the distal ileum (scale bar, 100 μm, n = 6). * P < 0.05, ** P < 0.01 by One-way ANOVA with post hoc Tukey's test (A, C–H, J). Data is presented as mean ± SEM.
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Cytotoxicity of TPP-45142 and its mechanism of action toward HER2-low breast cancer cell lines. A–D, TDCC of TPP-45142 for three T-cell donors compared with that of non-HER2 negative control (TPP-45161); co-cultures of human T cells with HCC1954, ZR-75-1, BT-20, or BT-549 cells were used at an E:T ratio of 5:1. E, TDCC of TPP-45142 for three T-cell donors compared with that of TPP-45161 in a co-culture of human T cells with BT20 3D spheroids at an E:T ratio of 1:5. F, T-cell activation induced by TPP-45142 as measured by expression of CD25 and CD69 expression on both CD4 + and CD8 + T cells as per FC analysis of ZR-75-1 and BT20 cells. G, Production of IFN-γ, IL2, IL6, IL8, IL10, and TNF-α cytokines in the culture supernatants obtained in the T-cell activation assay was measured using electrochemiluminescence assays.

Journal: Molecular Cancer Therapeutics

Article Title: TPP-45142—an Anti-HER2 T-cell Engager—Designed for Selective HER2-Low Cancer Immunotherapy

doi: 10.1158/1535-7163.MCT-25-0654

Figure Lengend Snippet: Cytotoxicity of TPP-45142 and its mechanism of action toward HER2-low breast cancer cell lines. A–D, TDCC of TPP-45142 for three T-cell donors compared with that of non-HER2 negative control (TPP-45161); co-cultures of human T cells with HCC1954, ZR-75-1, BT-20, or BT-549 cells were used at an E:T ratio of 5:1. E, TDCC of TPP-45142 for three T-cell donors compared with that of TPP-45161 in a co-culture of human T cells with BT20 3D spheroids at an E:T ratio of 1:5. F, T-cell activation induced by TPP-45142 as measured by expression of CD25 and CD69 expression on both CD4 + and CD8 + T cells as per FC analysis of ZR-75-1 and BT20 cells. G, Production of IFN-γ, IL2, IL6, IL8, IL10, and TNF-α cytokines in the culture supernatants obtained in the T-cell activation assay was measured using electrochemiluminescence assays.

Article Snippet: ZR-75-1 parental breast cancer tumor cells were purchased from the ATCC (ATCC CRL-1555) and cultured in complete medium, which included RPMI + GlutaMAX (Gibco, cat. #61870-036) supplemented with 10% heat-inactivated FBS (HI-FBS; Gibco-Invitrogen, cat. #10082-147) in 5% CO 2 at 37°C.

Techniques: Negative Control, Co-Culture Assay, Activation Assay, Expressing, Electrochemiluminescence

PK profiles and antitumor efficacy of TPP-45142 in the ZR-75-1 HER2-low breast cancer mouse model. A, TPP-45142 PK behavior in the ZR-75-1 xenograft model. Human T cells were administered to female NGS mice bearing intramammary ZR-75-1 tumors, and they were treated once with 89 Zr-TPP-45142 or the non-HER2 negative control 89 Zr-TPP-45161 ( n = 3). Microsamples (5 µL/time point) of blood were collected, and radioactivity was measured extemporaneously using a gamma counter (time: after radiolabeled-compound injection). B, Tumor accumulation of 89 Zr-TPP-45142 or non-HER2 negative control 89 Zr-TPP-45161 as measured by PET/CT imaging ( n = 3; time: after radiolabeled-compound injection). C, Antitumor activity of TPP-45142 in the ZR-75-1 xenograft model. Human T cells (10 × 10 6 ) were administered to female NSG mice bearing ZR-75-1 tumors, and they were treated on days 22 and 29 with TPP-45142 (500, 100, 50, and 10 μg/kg) and non-HER2 negative control TPP-45161 (500 μg/kg; n = 10 per group). ID, injected dose; MAD, median absolute deviation.

Journal: Molecular Cancer Therapeutics

Article Title: TPP-45142—an Anti-HER2 T-cell Engager—Designed for Selective HER2-Low Cancer Immunotherapy

doi: 10.1158/1535-7163.MCT-25-0654

Figure Lengend Snippet: PK profiles and antitumor efficacy of TPP-45142 in the ZR-75-1 HER2-low breast cancer mouse model. A, TPP-45142 PK behavior in the ZR-75-1 xenograft model. Human T cells were administered to female NGS mice bearing intramammary ZR-75-1 tumors, and they were treated once with 89 Zr-TPP-45142 or the non-HER2 negative control 89 Zr-TPP-45161 ( n = 3). Microsamples (5 µL/time point) of blood were collected, and radioactivity was measured extemporaneously using a gamma counter (time: after radiolabeled-compound injection). B, Tumor accumulation of 89 Zr-TPP-45142 or non-HER2 negative control 89 Zr-TPP-45161 as measured by PET/CT imaging ( n = 3; time: after radiolabeled-compound injection). C, Antitumor activity of TPP-45142 in the ZR-75-1 xenograft model. Human T cells (10 × 10 6 ) were administered to female NSG mice bearing ZR-75-1 tumors, and they were treated on days 22 and 29 with TPP-45142 (500, 100, 50, and 10 μg/kg) and non-HER2 negative control TPP-45161 (500 μg/kg; n = 10 per group). ID, injected dose; MAD, median absolute deviation.

Article Snippet: ZR-75-1 parental breast cancer tumor cells were purchased from the ATCC (ATCC CRL-1555) and cultured in complete medium, which included RPMI + GlutaMAX (Gibco, cat. #61870-036) supplemented with 10% heat-inactivated FBS (HI-FBS; Gibco-Invitrogen, cat. #10082-147) in 5% CO 2 at 37°C.

Techniques: Negative Control, Radioactivity, Injection, Positron Emission Tomography-Computed Tomography, Imaging, Activity Assay

FXR and HDCA regulates the levels of intestinal inflammation. (A) The relative expression of mRNA level in the distal ileum (n = 6). (B) Representative immunoblots of proteins in the distal ileum (n = 3). Relative expression levels of NLRP3 (C), IL-6 (D), TNF-α (E), IL-18 (F), IL-10 (G), IL-1β p17 (H). (I) Immunohistochemical staining and (J) quantitative analysis of NLRP3+, IL-6+, TNF-α+ and IL-18+ cells in the distal ileum (scale bar, 100 μm, n = 6). * P < 0.05, ** P < 0.01 by One-way ANOVA with post hoc Tukey's test (A, C–H, J). Data is presented as mean ± SEM.

Journal: Journal of Advanced Research

Article Title: Hyodeoxycholic acid relieves neuropathic pain by activating farnesoid X receptor signaling

doi: 10.1016/j.jare.2025.07.017

Figure Lengend Snippet: FXR and HDCA regulates the levels of intestinal inflammation. (A) The relative expression of mRNA level in the distal ileum (n = 6). (B) Representative immunoblots of proteins in the distal ileum (n = 3). Relative expression levels of NLRP3 (C), IL-6 (D), TNF-α (E), IL-18 (F), IL-10 (G), IL-1β p17 (H). (I) Immunohistochemical staining and (J) quantitative analysis of NLRP3+, IL-6+, TNF-α+ and IL-18+ cells in the distal ileum (scale bar, 100 μm, n = 6). * P < 0.05, ** P < 0.01 by One-way ANOVA with post hoc Tukey's test (A, C–H, J). Data is presented as mean ± SEM.

Article Snippet: Membranes were blocked using 5 % skim milk at room temperature for 1 h and subsequently incubated at 4 °C overnight with primary antibodies against FXR (1:1000, Immunoway, YN2161), zonula occludens-1 (ZO-1) (1:1000, Affinity, AF5145), Mucin-2 (1:1000, Abcam, ab272692), occludin (1:1000, Proteintech, 27260–1-AP), claudin-1 (1:1000, WL, WL03073), MMP-2 (1:1000, Immunoway, YT2798), MMP-9 (1:1000, Immunoway, YT1892), PPAR-γ (1:1000, Proteintech, 16643-1-AP), NOD-like receptor protein-3 (NLRP3) (1:1000, Immunoway, YT5382), interleukin-6 (IL-6) (1:1000, WL, WL02841), IL-18 (1:1000, WL, WL01127), tumor necrosis factor-alpha (TNF-α) (1:1000, WL, WL01581), IL-10 (1:1000, WL, WL03088), FGF15 (1:500, Santacruz, sc-514647), TGR5 (1:1000, Abcam, ab72608), BSEP (1:1000, Abcam, ab155421), CYP7α1 (1:1000, Abcam, ab65596) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:5000, Proteintech, 60004-1-Ig).

Techniques: Expressing, Western Blot, Immunohistochemical staining, Staining

HDCA regulates the inflammatory processes in the spinal cord. (A) Representative immunofluorescence staining results of IBA-1 in the spinal cord (scale, 100 μm, n = 3). (B) Immunofluorescence staining of CD86 in the spinal cord (scale, 100 μm, n = 3). (C) The mean fluorescence intensity of IBA-1 in spinal cord (n = 3). (D) The mean fluorescence intensity of CD86 in spinal cord (n = 3). (E) Relative expression of mRNA level in spinal cord (n = 6). (F) Representative immunoblots protein in spinal cord. (G) Relative expression levels of NLRP3, IL-10, IL-18, TNF-α protein (n = 3). * P < 0.05, ** P < 0.01by One-way ANOVA with post hoc Tukey's test (C, D, E, G). Data is presented as mean ± SEM.

Journal: Journal of Advanced Research

Article Title: Hyodeoxycholic acid relieves neuropathic pain by activating farnesoid X receptor signaling

doi: 10.1016/j.jare.2025.07.017

Figure Lengend Snippet: HDCA regulates the inflammatory processes in the spinal cord. (A) Representative immunofluorescence staining results of IBA-1 in the spinal cord (scale, 100 μm, n = 3). (B) Immunofluorescence staining of CD86 in the spinal cord (scale, 100 μm, n = 3). (C) The mean fluorescence intensity of IBA-1 in spinal cord (n = 3). (D) The mean fluorescence intensity of CD86 in spinal cord (n = 3). (E) Relative expression of mRNA level in spinal cord (n = 6). (F) Representative immunoblots protein in spinal cord. (G) Relative expression levels of NLRP3, IL-10, IL-18, TNF-α protein (n = 3). * P < 0.05, ** P < 0.01by One-way ANOVA with post hoc Tukey's test (C, D, E, G). Data is presented as mean ± SEM.

Article Snippet: Membranes were blocked using 5 % skim milk at room temperature for 1 h and subsequently incubated at 4 °C overnight with primary antibodies against FXR (1:1000, Immunoway, YN2161), zonula occludens-1 (ZO-1) (1:1000, Affinity, AF5145), Mucin-2 (1:1000, Abcam, ab272692), occludin (1:1000, Proteintech, 27260–1-AP), claudin-1 (1:1000, WL, WL03073), MMP-2 (1:1000, Immunoway, YT2798), MMP-9 (1:1000, Immunoway, YT1892), PPAR-γ (1:1000, Proteintech, 16643-1-AP), NOD-like receptor protein-3 (NLRP3) (1:1000, Immunoway, YT5382), interleukin-6 (IL-6) (1:1000, WL, WL02841), IL-18 (1:1000, WL, WL01127), tumor necrosis factor-alpha (TNF-α) (1:1000, WL, WL01581), IL-10 (1:1000, WL, WL03088), FGF15 (1:500, Santacruz, sc-514647), TGR5 (1:1000, Abcam, ab72608), BSEP (1:1000, Abcam, ab155421), CYP7α1 (1:1000, Abcam, ab65596) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:5000, Proteintech, 60004-1-Ig).

Techniques: Immunofluorescence, Staining, Fluorescence, Expressing, Western Blot