Figure 6 from Vitronectin Enrichment in Prostate Cancer Liver Metastases Promotes Adhesion and Survival
Year: 2026
Type: other
Abstract: The adhesion effects of VTN are robust across assay conditions and biological sources. A, Representative microscopy of prostate cancer cells plated on polyHEMA-coated plates depicting the abrogation of cell attachment and successful spheroid formation. B, Adhesion assays were performed with polyHEMA-coated plates and serum-free media with or without 5 μg/mL VTN. Bars show three biological replicates, and significance was calculated with two-tailed Welch t tests. ns, not statistically significant. C, Cells were subjected to a time-course adhesion assay in serum-free media with or without 5 μg/mL VTN. Bars show three biological replicates, and significance was calculated for the 24-hour timepoint with two-tailed Welch t tests. D, Images were taken of well centers from the 24-hour timepoint, and cells within captured fields of view were counted. Results are for three biological replicates, and two-tailed Welch t tests were used for statistical determinations. E, Adhesion assays were performed in serum-free media with 5 μg/mL rVTN, 5 μg/mL human plasma–purified VTN (hVTN), or no VTN. Complete, use of complete media as a positive control. Bars show three biological replicates, and significance was calculated with two-tailed Welch t tests. F, Conditioned media was recovered from 5-donor human plateable hepatocytes (first collection period/day 1). An adhesion assay was performed with 22Rv1 and 100% conditioned media serially diluted 1:1 for seven timepoints. Images show representative regions of adhered or suspended 22Rv1 cells following exposure to 100%, 50%, 25%, or 0% conditioned media. G, Conditioned media (24 μL) was directly mixed with 8 μL of 4× SDS-PAGE and immunoblotted for VTN as specified in “Materials and Methods.” As media was collected and replenished every 24 hours, lane labels indicate the period of media collection since hepatocyte plating (day 1 corresponds to collection on day 1 since plating). Uncond. media, unconditioned hepatocyte maintenance media. H, An adhesion assay was performed with basal hepatocyte maintenance media, 50% hepatocyte-conditioned maintenance media, or complete culture media as a positive control (RPMI-1640 with 10% FBS). Complete, use of complete culture media as a positive control; maintenance, use of maintenance media as the basal serum-free media (rather than RPMI-1640). Bars show three biological replicates, and significance was calculated with two-tailed Welch t tests. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001.
Subfield: Immunology and Allergy
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W7213460570
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