Journal: International Journal of Molecular Sciences

Article Title: Tedizolid Targets AQP9-JAK/STAT Axis to Suppress Metastatic Progression in Clear Cell Renal Cell Carcinoma: Mechanism and Therapeutic Implications

doi: 10.3390/ijms27104234

Figure Lengend Snippet: AQP9 activates the JAK/STAT signaling pathway in ccRCC cells. ( A ) A KEGG pathway enrichment analysis of genes co-expressed with AQP9. ( B ) A GSEA plot showing significant enrichment of the JAK/STAT signaling pathway gene set in tumors with high AQP9 expression. ( C , D ) A Western blot analysis of key JAK/STAT pathway proteins (JAK2, STAT3, and their phosphorylated forms p-JAK2 and p-STAT3) in Caki-1 and 786-O cells after AQP9 knockdown or overexpression. β-actin served as a loading control. The data are presented as mean ± SD. **** p < 0.0001.

Article Snippet: The samples were resolved by SDS-PAGE, transferred onto PVDF membranes, and then incubated overnight at 4 °C with primary antibodies against AQP9 (sc-74409 G-3, 1:100, Santa Cruz Biotechnology, Dallas, TX, USA)), N-cadherin (A0433, 1:1000, ABclonal, Wuhan, China), E-cadherin (A0433, 1:1000, ABclonal, China), Vimentin (10366-1-AP, 1:20,000, Proteintech, Rosemont, IL, USA), Snail (A5243, 1:1000, ABclonal, China), MMP2 (A19080, 1:1000, ABclonal, China), MMP9 (F0008, 1:1000, Selleck Chemicals, Houston, TX, USA), phosphorylated JAK2 (AP0531, 1:1000, ABclonal, China), JAK2 (K5N8, 1:1000, Selleck, WA, USA), phosphorylated STAT3 (B17F7, 1:2000, Selleck, WA, USA), STAT3 (WL01836, 1:1000, Wanleibio, Shenyang, China), and β-actin (AC038, 1:20,000, ABclonal, China).

Techniques: Expressing, Western Blot, Knockdown, Over Expression, Control

Journal: International Journal of Molecular Sciences

Article Title: Tedizolid Targets AQP9-JAK/STAT Axis to Suppress Metastatic Progression in Clear Cell Renal Cell Carcinoma: Mechanism and Therapeutic Implications

doi: 10.3390/ijms27104234

Figure Lengend Snippet: AQP9 regulates EMT in ccRCC via the JAK/STAT pathway. ( A , C ) A wound healing assay showing that the STAT3 inhibitor Stattic attenuates the enhanced migration of ccRCC cells induced by AQP9 overexpression. ( B , D ) Transwell migration and invasion assays showing that Stattic attenuates the enhanced migration and invasion of ccRCC cells induced by AQP9 overexpression. ( E ) A Western blot analysis showing that Stattic reverses the activation of the JAK2/STAT3 pathway induced by AQP9 overexpression. ( F ) A Western blot analysis showing that Stattic reverses the promotion of EMT markers induced by AQP9 overexpression. The data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The samples were resolved by SDS-PAGE, transferred onto PVDF membranes, and then incubated overnight at 4 °C with primary antibodies against AQP9 (sc-74409 G-3, 1:100, Santa Cruz Biotechnology, Dallas, TX, USA)), N-cadherin (A0433, 1:1000, ABclonal, Wuhan, China), E-cadherin (A0433, 1:1000, ABclonal, China), Vimentin (10366-1-AP, 1:20,000, Proteintech, Rosemont, IL, USA), Snail (A5243, 1:1000, ABclonal, China), MMP2 (A19080, 1:1000, ABclonal, China), MMP9 (F0008, 1:1000, Selleck Chemicals, Houston, TX, USA), phosphorylated JAK2 (AP0531, 1:1000, ABclonal, China), JAK2 (K5N8, 1:1000, Selleck, WA, USA), phosphorylated STAT3 (B17F7, 1:2000, Selleck, WA, USA), STAT3 (WL01836, 1:1000, Wanleibio, Shenyang, China), and β-actin (AC038, 1:20,000, ABclonal, China).

Techniques: Wound Healing Assay, Migration, Over Expression, Western Blot, Activation Assay