Fig 1: FN-binding integrins mediate ECM production. Knockdown of FN or FN-binding integrins in CAFs prevents the formation of FN and COL fibers. CAF-1299 cells were treated with siRNA for a scramble control vs. FN, ITGA5, or ITGB3. After 72 hours, IF staining shows FN and COL content. The top set of images were not permeabilized. Images are representative of at least three independent experiments. Blots confirm knockdown. si-CTRL, CAFs with siRNA for a Universal Negative Control; si-FN, CAFs with siRNA-mediated knockdown of FN1; si-α5, CAFs with ITGA5 knockdown; si-β3, CAFs with ITGB3 knockdown.
Fig 2: Tumor cells utilize CAF-produced FN to overcome isolation stress. A, CAFs enhance tumor initiation via FN (orthotopic pancreas cancer model). Luciferase-expressing KP4 PDAC cells were orthotopically injected (with or without CAF-1299) into the pancreas of nu/nu mice. After 3 weeks, luciferase imaging was performed to survey tumor establishment in the pancreas. B, FN knockdown prevents CAF assembly of COL fibers. CAF-1299 and CAF-1424 cells were treated with siRNA for a scramble control vs. FN. After 72 hours, IF staining shows FN and COL content. Images are representative of at least three independent experiments. C, Knockdown of FN in CAFs increases soluble COL in media. CAF-1299 cells were cultured for 72 hours, and then the conditioned media and cell lysates were collected and processed for immunoblotting to confirm FN knockdown and assess the level of soluble COL secreted by the cells into the media. si-CTRL, siRNA Universal Negative Control; si-FN, siRNA-mediated knockdown of FN1.
Supplier Page from Novus Biologicals, a Bio-Techne Brand for Fibronectin Antibody (DH1)
Available conjugates: Available conjugates: UnconjugatedSpecificity: The antibody is specific to extradomain A (EDA) sequence of a cellular fibronectin and recognizes thus only the cellular fibronectinSizes Available: 0.025 mg