2.

HS promotes CRC proliferation in vitro. (A) CRC cell lines were exposed to HS at the temperatures indicated for 10 minutes, followed by cell culture in complete medium for 48 hours. Cells were double labeled with Alexa fluor 488-Annexin V and PI, then analyzed by FACS. The results shown are representative from three independent experiments. (B, C) Proliferation of CT-26 and SW620 cells treated by different temperatures’ HS as measured by BrdU incorporation. BrdU incorporation was detected by immunofluorescent staining (B, C). The percentage of BrdU positive cells (BPI) in CT-26 cells (B) and SW620 cells (C) were quantitatively measured. Red, BrdU; Green, phalloidin; Blue, DAPI (200 ×). P values were determined by one-way ANOVA test. * P < 0.05, ** P < 0.01 ( n = 3).