Fig. 6
Effect of DAC treatment on the expression of the NY-ESO-1 tumor antigen in CRC 1076 Col and 1872 Col cell lines. (A, B) RT-qPCR analysis showed an induction of NY-ESO-1 in both 1076 Col and 1872 Col after 5 μΜ of DAC treatment. (C, D) Western blot analysis showed a significant induction of NY-ESO-1 in 1872 (fivefold, ****p = 0.0001) than in 1076 (1.4-fold, ****p = 0.0001) Col following treatment with DAC. (E, F) Confocal microscopy images showing the localization of NY-ESO-1 (white arrows) in 1076 Col and 1872 Col cells treated with 5 μM of DAC compared to the untreated cells. Cells were treated with DAC for 48 h. and then stained with Alexa-fluor labeled 12D7-conjugated antibody as described in the methods section. Fluorescence images were captured at 60X magnification. Arrows presented in the confocal images indicate that induced NY-ESO-1 is located at the nucleus. The data are from duplicate experiments and presented as mean ± SD. Scale bar is 10 μm in (E). Scale bar is 20 μm