A CLIC1 network coordinates matrix stiffness and the Warburg effect to promote tumor growth in pancreatic cancer. Zheng et al.

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In a study exploring the intricate relationship between PDAC matrix stiffness and the Warburg effect, Zheng et al. reveal CLIC1 as a critical mediator. Matrix stiffness increases CLIC1 levels through the Wnt/β-catenin/TCF4 pathway, correlating with adverse outcomes and tumor progression. Reducing matrix stiffness may offer therapeutic benefits by targeting CLIC1. This data is the original immunoblotting data of the above study.

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Western blotting analysis Cells were lysed using the RIPA Lysis and Extraction Buffer (Thermo Scientific, #89901) containing phosphatase and protease inhibitors. Protein concentration was quantified using the BCA Protein Assay Kit (ShareBio, #SB-WB013). Subsequently, proteins were separated by SDS-PAGE and transferred onto nitrocellulose membranes (Millipore, Danvers, MA). Non-specific binding sites on the membrane were blocked at room temperature for 1 hour with Tris-buffered saline with Tween20 (TBST) containing 5% non-fat dry milk. The membranes were incubated overnight at 4°C with one of the following primary antibodies: anti-CLIC1 (1:1000), anti-HIF1α (1:1000), anti-HIF1α-OH (1:1000), anti-TCF4 (1:1000), anti-SPI1 (1:1000), anti-FOXA2 (1:1000), anti-EGR2 (1:1000), anti-β-catenin (1:1000), anti-HK2 (1:1000), anti-GLUT1 (1:1000), anti-LDHA (1:1000), anti-β-actin (1:5000) and anti-Histone-H3 (1:3000). Following three washes with TBST, the membranes were incubated with species-specific secondary antibodies. The bound secondary antibodies were detected by a chemiluminescence fluorescence imaging system (BIO-RAD, ChemiDoc XRS+, 1708265).

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