Fig. 2

Targeting Validation of functionalized multi-color nanoprobes in vitro. Western blots indicate the relative expression levels of three different cancer biomarkers: a The 4175TR cell line shows increased CAV1 receptor expression relative to the MCF7 cell line. b MCF7 cells show higher CXCR4 expression than 4175TR cells. c SKOV3 ovarian cancer cell line used in this study were determined to be FR positive by immunoblotting. The western blots in a, b an c panels have been cropped to highlight the protein of interest in each panel. Panel a, b and c show western blots of caveolin-1, CXCR4 and folate receptor- alpha respectively. The full-length western blots are presented in Supplementary Fig. 9. d Flow cytometry shows a statistically significant increase in uptake of CAV1-targeted nanoprobes (Rh-fHoANCs) compared to untargeted nanoprobes (Rh-HoANCs) in CAV1-expressing 4175TR cells. No significant increase was observed in uptake of CAV1-targeted versus untargeted nanoprobes in CAV1-negative MCF7 cells. e CXCR4-targeted nanoprobes (FITC-fErANCs) showed significantly increased uptake in CXCR4-expressing MCF7 cells compared to untargeted nanoprobes (FITC-ErANCs). No significant increase was observed in uptake of CXCR4-targeted versus untargeted nanoprobes in CXCR4-negative 4175TR cells. f A statistically significant increase in uptake of FRα-targeted nanoprobes (647-fTmANCs) compared to untargeted nanoprobes (647-TmANCs) was observed in FRα-expressing SKOV3 cells. For panel d, e and f student t-test was used to determine statistical significance between two groups