Fig 1: Expression of HDAC2 and heterochromatin proteins after cisplatin incubationA. Western blots for HDAC2, HP1α, HP1β, and HP1γ in PEO1 and PEO4 cells with or without cisplatin treatment (6 uM, 24h). Membranes were probed with the indicated antibodies, and tubulin was used as a loading control. B. HDAC2 and HP1 protein expression detected by immunofluorescence. Cells were seeded on cover slips and fixed as described in the Materials and Methods in PEO1 cells after cisplatin treatment for 6h and 24h. Alexa488 (green channel) and DAPI (blue channel) were used to stain target proteins and the nuclei, respectively. Images were taken using a confocal microscope. C. Expression of HDAC2, HP1α, HP1β, and HP1γ mRNA in PEO1 cells measured by RT-PCR as described in the Materials and Methods. Relative expression of the target gene was calculated as the average ΔCt and normalized to that of the housekeeping gene β-actin. Results are as presented as mean ±SD from biological triplicates. *p<0.05, **p<0.01 (Student's t-test).