Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Baicalin Protects Mouse Testis From Injury Induced by PM 2.5 Associated With the DKK1‐Wnt/β‐Catenin Signaling Pathway

doi: 10.1002/jbt.71057

Figure Lengend Snippet: Effect of baicalin on testosterone level and gene expression of testosterone synthesis pathway in mice exposed to PM 2.5 . (A–C) Baicalin increased the expression of StAR, CYP11A1, and CYP17A1 mRNA in testis of mice exposed to PM 2.5 . (D) Effect of baicalin on testosterone content in serum of mice exposed to PM 2.5 . (E) StAR, CYP11A1, CYP17A1, and β‐actin representative protein band. (F–H) Effect of baicalin on StAR, CYP11A1, and CYP17A1 protein expression. The data are expressed as mean ± SD ( n = 6 per group). * p < 0.05 and ** p < 0.001, compared with the control group; # p < 0.05 and ## p < 0.001, compared with PM 2.5 exposure group.

Article Snippet: Antibodies against DKK1, Wnt4 LRP5/6, β‐catenin, StAR, CYP17A1, CYP11A1, and β‐actin were all purchased from ABclonal Trading Co. Ltd. (Shanghai, China), Proteintech Group Inc. (Wuhan, China), and Abcam plc (Cambridge, USA).

Techniques: Gene Expression, Expressing, Control

Journal: bioRxiv

Article Title: Mast Cell Specific Cyp11a1 Deficiency Promotes T Cell Mediated Immunity and Suppresses Tumour Metastasis in a Mouse Model of Melanoma

doi: 10.64898/2026.03.04.709617

Figure Lengend Snippet: ( a ) Representative images of lung tumour nodules from control (Cyp11a1-sufficient) and mast cell–specific Cyp11a1 knockout (cKO) mice following metastatic challenge. Note the greater tumour burden (black arrows) in control vs. cKO animals. ( b ) Quantification of lung metastatic nodules. ( c ) Body weight monitoring of mice during metastatic progression. ( d ) Histological (H&E) and toluidine blue staining showing tumour tissue morphology with enhanced nodule formation (red arrows) and greater mast cell infiltration (black arrows) in lung tissues from control (Cyp11a1-sufficient) mice compared to mast cell–specific Cyp11a1 knockout mice. ( e ) Quantitative assessment of mast cells in lungs of control- and cKO mice (at least three-five high-power fields per section were counted). (f-g) Quantitative RT-PCR (qPCR) analysis of the expression of mast cell marker genes ( Mcpt1 and Mcpt6 ) in lungs of tumour-bearing control (Ctrl) and cKO mice. Gene expression was normalized to GAPDH and Actb using the ΔΔCt method and presented as fold change. Statistical analysis was performed using unpaired t -tests (For tumour nodule enumeration, n = 8; histology, n = 3; qPCR, n = 4). P < 0.05 was considered significant.

Article Snippet: Slides were then incubated overnight at 4°C in the dark with the following primary antibodies: Monoclonal Mouse Mast Cell Tryptase (ab2378, Abcam) diluted 1:100, CYP11A1 (D8F4F) Rabbit mAb (Cell Signaling Technology, #14217) diluted 1:200, Mouse CD45 (FITC) diluted 1:200, and CD107a PE diluted 1:200.

Techniques: Control, Knock-Out, Staining, Quantitative RT-PCR, Expressing, Marker, Gene Expression

Journal: bioRxiv

Article Title: Mast Cell Specific Cyp11a1 Deficiency Promotes T Cell Mediated Immunity and Suppresses Tumour Metastasis in a Mouse Model of Melanoma

doi: 10.64898/2026.03.04.709617

Figure Lengend Snippet: ( a ) Confocal microscopy images showing localization of the mast cell marker tryptase (red) and Cyp11a1 (green) in lung tumours from control (Ctrl; Cyp11a1-sufficient) and mast cell–specific Cyp11a1 knockout (cKO) mice. ( b ) Quantitative analysis of Cyp11a1/tryptase co-localization using ImageJ software confirming a significantly higher proportion of Cyp11a1□ mast cells in control tumours compared to cKO tumours ( c ) Quantitative RT-PCR (qPCR) analysis of Cyp11a1 gene expression in lungs of tumour-bearing control (Ctrl) and cKO mice. Cyp11a1 gene expression was normalized to Gapdh and Actb expression using the ΔΔCt method and presented as fold change. Statistical analysis was performed using unpaired t-tests (For IF staining, n = 3; for qPCR, n= 4). P < 0.05 was considered significant.

Article Snippet: Slides were then incubated overnight at 4°C in the dark with the following primary antibodies: Monoclonal Mouse Mast Cell Tryptase (ab2378, Abcam) diluted 1:100, CYP11A1 (D8F4F) Rabbit mAb (Cell Signaling Technology, #14217) diluted 1:200, Mouse CD45 (FITC) diluted 1:200, and CD107a PE diluted 1:200.

Techniques: Confocal Microscopy, Marker, Control, Knock-Out, Software, Quantitative RT-PCR, Gene Expression, Expressing, Staining

Journal: bioRxiv

Article Title: Mast Cell Specific Cyp11a1 Deficiency Promotes T Cell Mediated Immunity and Suppresses Tumour Metastasis in a Mouse Model of Melanoma

doi: 10.64898/2026.03.04.709617

Figure Lengend Snippet: ( a, c ) Confocal microscopy images showing expression of the general leukocyte marker CD45 (red), the activation marker CD107a (magenta) and Cyp11a1 in lung cells isolated from control (Cyp11a1-sufficient) and mast cell–specific Cyp11a1 knockout (cKO) tumour-bearing mice. ( b, d ) Quantitative analysis of the intensity of Cyp11a1, CD45 and CD107a staining intensity in lung tissue cells from control mice (Ctrl) and mice with mast cell-specific depletion of Cyp11a1 (cKO), using the ImageJ software. VoF, view of field. Data represent mean values ±□SEM; P □<□0.05 by unpaired two-tailed t -test (n = 3).

Article Snippet: Slides were then incubated overnight at 4°C in the dark with the following primary antibodies: Monoclonal Mouse Mast Cell Tryptase (ab2378, Abcam) diluted 1:100, CYP11A1 (D8F4F) Rabbit mAb (Cell Signaling Technology, #14217) diluted 1:200, Mouse CD45 (FITC) diluted 1:200, and CD107a PE diluted 1:200.

Techniques: Confocal Microscopy, Expressing, Marker, Activation Assay, Isolation, Control, Knock-Out, Staining, Software, Two Tailed Test

Journal: bioRxiv

Article Title: Mast Cell Specific Cyp11a1 Deficiency Promotes T Cell Mediated Immunity and Suppresses Tumour Metastasis in a Mouse Model of Melanoma

doi: 10.64898/2026.03.04.709617

Figure Lengend Snippet: Immunophenotyping of lung tumours by flow cytometry. Note that no significant effect of mast cell-specific depletion of Cyp11a1 on the proportion of T cells, B cells, NK cells, eosinophils or basophils among the CD45 + population were seen (gating strategy shown in Suppl. Fig. 1 ). Data represent mean□values ±□SEM; P □<□0.05 by unpaired two-tailed t -test (Ctrl, n = 8; cKO, n = 7 - 8).

Article Snippet: Slides were then incubated overnight at 4°C in the dark with the following primary antibodies: Monoclonal Mouse Mast Cell Tryptase (ab2378, Abcam) diluted 1:100, CYP11A1 (D8F4F) Rabbit mAb (Cell Signaling Technology, #14217) diluted 1:200, Mouse CD45 (FITC) diluted 1:200, and CD107a PE diluted 1:200.

Techniques: Flow Cytometry, Two Tailed Test

Journal: bioRxiv

Article Title: Mast Cell Specific Cyp11a1 Deficiency Promotes T Cell Mediated Immunity and Suppresses Tumour Metastasis in a Mouse Model of Melanoma

doi: 10.64898/2026.03.04.709617

Figure Lengend Snippet: Total dispersed lung cell populations were prepared from tumour-bearing control mice (Ctrl) and mice with Cyp11a1 -deficient mast cells (cKO), and were activated by incubation with PMA. ( a ) After activation of cells, the expression of IFN-γ and TNFα in CD8 + T cells, CD4 + T cells, NK cells, B cells and mast cells was assessed by flow cytometry as indicated. ( b ) qPCR analysis of IFN-γ and TNFα gene expression in lungs from tumour-bearing control mice (Ctrl) and mice with Cyp11a1 -deficient mast cells (cKO). Data represent mean□±□SEM; P □<□0.05 by unpaired two-tailed t -test (at least n = 6/group).

Article Snippet: Slides were then incubated overnight at 4°C in the dark with the following primary antibodies: Monoclonal Mouse Mast Cell Tryptase (ab2378, Abcam) diluted 1:100, CYP11A1 (D8F4F) Rabbit mAb (Cell Signaling Technology, #14217) diluted 1:200, Mouse CD45 (FITC) diluted 1:200, and CD107a PE diluted 1:200.

Techniques: Control, Incubation, Activation Assay, Expressing, Flow Cytometry, Gene Expression, Two Tailed Test