Figure 2.

Exposure of HEY cells to paclitaxel enhances phosphorylation of Src in a time dependent manner. (A) The expression of p-Src and t-Src was assessed by immunofluorescence in untreated and paclitaxel (0.05 µg/mL) treated cells following 6, 24 or 72 h of incubation. Staining was visualized using the secondary Alexa 590 (red) fluorescent-labelled antibody, and nuclei were detected by DAPI (blue) staining. Magnification 400× scale bar = 250 µm. Quantification of (B) p-Src and (C) t-Src fluorescent intensities were conducted using Fiji software. Results are displayed as the percentage of the average fluorescent intensity relative to untreated cells ± SEM (n = 3/group). (D) Total cell lysates of HEY cells were collected at 6, 24 and 72 h after paclitaxel treatment and were subjected to immunoblot analysis using antibodies specific for p- or t-Src or GAPDH. Images are representative of three independent experiments. Densitometry analysis of (E) p-Src and (F) t-Src protein expressions. The values represent the relative mean of band intensity normalized to GAPDH loading control ± SEM. Significance is indicated by * p < 0.05, ** p < 0.01.