jak2 phosphorylation  (MedChemExpress)

Journal: Animal Bioscience

Article Title: ELF5 modulates casein synthesis in goat mammary epithelial cells via JAK2/STAT5 signaling pathway

doi: 10.5713/ab.25.0181

Figure Lengend Snippet: ELF5 regulate casein synthesis in GMECs. (A, B) The expression of αS1-casein, αS2-casein, β-casein, κ-casein, phosphorylated JAK2 and STAT5 after transfection with pcDNA3.1-ELF5 or pcDNA3.1-NC for 48 h. (C, D) The expression of αS1-casein, αS2-casein, β-casein, κ-casein, p-JAK2, and p-STAT5 after transfection with siRNA-ELF5 or siRNA-NC (100 nM) for 48 h. The relative protein abundance of αS1-casein, αS2-casein, β-casein, and κ-casein was normalized to β-tubulin. The relative protein abundance of p-JAK2 and p-STAT5 was normalized to total JAK2 and STAT5, respectively. Values are presented as mean±SEM. * p<0.05, ** p<0.01. siRNA, small interfering ribonucleic acid; NC, negative control; GMECs, goat mammary epithelial cells; SEM, standard error of the mean.

Article Snippet: For the inhibition of JAK2/STAT5 pathway, JAK2 inhibitor AG490 (30 μM, MedChemExpress) or STAT5 inhibitor STAT5-IN-1 (50 μM, MedChemExpress) were treated with cells.

Techniques: Expressing, Transfection, Quantitative Proteomics, Negative Control

Journal: Animal Bioscience

Article Title: ELF5 modulates casein synthesis in goat mammary epithelial cells via JAK2/STAT5 signaling pathway

doi: 10.5713/ab.25.0181

Figure Lengend Snippet: The protein abundance of caseins after JAK2 and STAT5 inhibition in GMECs. (A) The expression of αS1-casein, αS2-casein, β-casein, κ-casein, and phosphorylated JAK2 and STAT5 after JAK2 inhibitor Tyrphosting AG490 (30 μM) treatment for 48 h. (B) The expression of αS1-casein, αS2-casein, β-casein, κ-casein, and p-STAT5 after STAT5 inhibitor STAT5-IN-1 (50 μM) treatment for 48 h. The relative protein abundance of αS1-casein, αS2-casein, β-casein, and κ-casein was normalized to β-tubulin. The relative protein abundance of p-JAK2 and p-STAT5 was normalized to total JAK2 and STAT5, respectively. Values are presented as mean±SEM. ** p<0.01. GMECs, goat mammary epithelial cells; SEM, standard error of the mean.

Article Snippet: For the inhibition of JAK2/STAT5 pathway, JAK2 inhibitor AG490 (30 μM, MedChemExpress) or STAT5 inhibitor STAT5-IN-1 (50 μM, MedChemExpress) were treated with cells.

Techniques: Quantitative Proteomics, Inhibition, Expressing

Journal: Animal Bioscience

Article Title: ELF5 modulates casein synthesis in goat mammary epithelial cells via JAK2/STAT5 signaling pathway

doi: 10.5713/ab.25.0181

Figure Lengend Snippet: ELF5 mediates casein synthesis by JAK2 and STAT5 activity in GMECs. (A) Cells were treated with Tyrphosting AG490 (30 μM) or DMSO, followed by pcDNA3.1-ELF5 or pcDNA3.1-NC transfection for 48 h. (B) Cells were treated with Tyrphosting AG490 (30 μM) or DMSO, followed by siRNA-ELF5 or siRNA-NC (100 nM) transfection for 48 h. The protein abundances of αS1-casein, αS2-casein, β-casein, κ-casein, p-JAK2, and p-STAT5 were detected. The relative protein abundance of αS1-casein, αS2-casein, β-casein, and κ-casein was normalized to β-tubulin. The relative protein abundance of p-JAK2 and p-STAT5 was normalized to total JAK2 and STAT5, respectively. Values are presented as mean±SEM. a–d Different lowercase letters represent significant differences (* p<0.05). NC, negative control; siRNA, small interfering ribonucleic acid; GMECs, goat mammary epithelial cells; SEM, standard error of the mean.

Article Snippet: For the inhibition of JAK2/STAT5 pathway, JAK2 inhibitor AG490 (30 μM, MedChemExpress) or STAT5 inhibitor STAT5-IN-1 (50 μM, MedChemExpress) were treated with cells.

Techniques: Activity Assay, Transfection, Quantitative Proteomics, Negative Control

Journal: Animal Bioscience

Article Title: ELF5 modulates casein synthesis in goat mammary epithelial cells via JAK2/STAT5 signaling pathway

doi: 10.5713/ab.25.0181

Figure Lengend Snippet: Molecular mechanism of ELF5 promotes casein synthesis by enhancing the activity of JAK2/STAT5 signaling pathway in goat mammary epithelial cells. PRLR stands for prolactin receptor.

Article Snippet: For the inhibition of JAK2/STAT5 pathway, JAK2 inhibitor AG490 (30 μM, MedChemExpress) or STAT5 inhibitor STAT5-IN-1 (50 μM, MedChemExpress) were treated with cells.

Techniques: Activity Assay

Journal: Breast Cancer Research : BCR

Article Title: Modulating phosphodiesterase-5 activity to suppress the immunosuppressive mechanisms of myeloid-derived suppressor cells in breast cancer

doi: 10.1186/s13058-025-02131-5

Figure Lengend Snippet: Tadalafil disrupted the immunosuppressive ability of MDSCs by inhibiting STAT3 transcription. A RT-qPCR analysis of the expression of the BM-MDSCs stat3 . B Western blot analysis was used to detect STAT3 expression in BM-MDSCs after tadalafil treatment. C Flow cytometry representative plot of the effect of tadalafil treatment combined with administration of the STAT3 inhibitor on the expression of arginase 1, a functional marker of MDSCs, induced in vitro. D Transwell co-culture plate design. E Representative images and quantification of heme staining in transwell chambers. Data represents the mean ± SD ( n ≥ 3), **P < 0.01 and **P < 0.001 versus vehicle

Article Snippet: Tadalafil (HY-90009 A), STAT3 inhibitor (Tyrphostin AG490, HY-12000) and PARP-1 inhibitor (AG14361, HY-12032) were purchased from MedChem Express (New Jersey, USA).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Flow Cytometry, Functional Assay, Marker, In Vitro, Co-Culture Assay, Staining

Journal: Breast Cancer Research : BCR

Article Title: Modulating phosphodiesterase-5 activity to suppress the immunosuppressive mechanisms of myeloid-derived suppressor cells in breast cancer

doi: 10.1186/s13058-025-02131-5

Figure Lengend Snippet: Tadalafil regulates STAT3 transcription through PARP1. A GSEA analysis revealed significant alterations in NAD-related signaling pathways following tadalafil intervention in BM-MDSCs. B Heatmap displaying the expression patterns of genes associated with NAD-related signaling pathways. C Flow cytometry gating strategy and analysis of PARP1 median fluorescence intensity in BM-MDSCs. D Western blot analysis of cGKI and PARP1 expression in MDSC cells after tadalafil treatment. E The expression level of PARP1 mRNA in BM-MDSCs. F Immunofluorescence staining assessing PARP1 expression in BM-MDSCs post-tadalafil treatment. G Western blot detection of PARP1, STAT3, and phosphorylated STAT3 (p-STAT3) levels in cytoplasmic and nuclear fractions of BM-MDSCs upon tadalafil intervention. H Western blot analysis of STAT3 and p-STAT3 expression changes in BM-MDSCs treated with tadalafil in the presence of a PARP1 inhibitor. I Flow cytometry gating strategy and analysis of p-STAT3 median fluorescence intensity in BM-MDSCs. J Genotyping verification of PARP1 knockout mice. K Western blot confirmation of PARP1 knockout efficiency. L Western blot evaluation of Arg1, STAT3, and p-STAT3 expression in BM-MDSCs from PARP1 knockout mice following tadalafil intervention. Data represents the mean ± SD ( n ≥ 3), *P < 0.05, **P < 0.01 and **P < 0.001 vs. vehicle

Article Snippet: Tadalafil (HY-90009 A), STAT3 inhibitor (Tyrphostin AG490, HY-12000) and PARP-1 inhibitor (AG14361, HY-12032) were purchased from MedChem Express (New Jersey, USA).

Techniques: Protein-Protein interactions, Expressing, Flow Cytometry, Fluorescence, Western Blot, Immunofluorescence, Staining, Knock-Out