Supplementary Figures 1 - 3 from Relocation of CLIC1 Promotes Tumor Cell Invasion and Colonization of Fibrin
Year: 2023
Type: supplementary-materials
Abstract: Supplementary Fig. S1 (A-B), western blotting for CLIC1 to demonstrate knockdown in shCLIC1 786-0 and HT1080 clones compared to shSCR control (A) as well as in HUVEC cells compared to non-targeted control siRNA (B). For both cell types, clones shCLIC1-2 and shCLIC1-4 were selected for further experiments. (C), fluorescence microscopy images of fibrin clots were analyzed for percent propidium iodide-positive 786-0 per optical field after CLIC1 silencing with two CLIC1 shRNA vectors (shCLIC1-2 and 4) compared to scrambled shRNA (shSCR). Not significant (ns), p > 0.05. Supplementary Fig. 2 (A), CLIC1 fluorescence intensity is depicted as a ratio of peripheral to interior CLIC1 in shβ3-786-0 cells compared to shSCR-786-0 cells to assess CLIC1 redistribtion to invadopodia at indicated times. ***P < 0.001, *P <0.05. (B-C), western blotting for β3 integrin expression in shSCR-786-0 compared to shβ3 (B) and shCLIC1 (C). Ponceau S staining shows equal loading. Supplementary Fig. S3 (A), western blotting for RhoA-GTP in shSCR and shCLIC1 786-0 cells following rhotekin-RBD pull-down using the RhoA activation assay biochem kit (Cytoskeleton). (B), 786-0 cells transformed with shSCR (left) or shCLIC1 (right) cultured in 3D fibrin for 48 hours were fixed in 4% PFA, then washed in PBS, blocked in 2% BSA and probed for fibronectin (green) prior to confocal microscopy. Nuclei are stained with draq5 (blue).
Language: English
Subfield: Immunology and Allergy
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W4362529334