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Figure 4 | BMC Cancer

Figure 4

From: Mammary tumors that become independent of the type I insulin-like growth factor receptor express elevated levels of platelet-derived growth factor receptors

Figure 4

Downregulation of PDGFRs in IGF-IR-independent recurrent cells increased cell number. MTT assays were performed to examine cell proliferation/survival after siRNA-mediated knockdown of PDGFRα and β alone or in combination (a). Cell number is expressed relative to transfection with the control oligonucleotide and represents the average of five trials. JC1 assays were used to assess apoptosis. Cells were plated in 12-well dishes and transfected with siRNA oligonucleotides specific to PDGFR. JC1 was added after 48 h and fluorescent green cells were quantified. Results represent the average of 3-4 trials relative to controls (b). Apoptosis was also evaluated by immunofluorescence staining for cleaved Caspase-3 (c). Proliferation was quantified using immunofluorescence staining for Ki67 (d) and phosphorylated HistoneH3 (e). For all three above assays, cells were plated in 6-well dishes on coverslips and transfected as above. Fixing and staining was performed 48 h post transfection. Images were collected at 100 × magnification and positive nuclei were counted. Results represent the average percent positivity relative to controls for three individual trials (d, e, f). * = p < 0.05 (relative to respective control).

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