Figure 4

HSP60 protein decreased the proliferation rate and colony formation ability of PcDNA3.1( IGFBP7 )-RKO cells. A: PcDNA3.1(IGFBP7)-RKO cells were plated in sextuple in 96-well microtitre plates at 3 × 10³/well, cultured with medium with or without recombinant HSP60(1 μg/ml). Ten μl of CCK8 was added to each well at the indicated time (12 h, 24 h, 36 h, 48 h, 60 h, 72 h). Cellular viability was determined by measuring the absorbance of the converted dye at 450 nm two hours after adding CCK8. Values shown are mean ± SD for sextuple cultures from one experiment, representive of three independent experiments conducted. B: PcDNA3.1(IGFBP7)-RKO cells (500/well) were seeded into 0.3% Bacto-agar over a 0.6% agar bottom layer in triplicate in 6-well plates, with or without 1 μg/ml HSP60. After 3 weeks of incubation, colony number (>100 μm)were analyzed. Values are mean ± S.D for data from three independent experiments. C: Colony size was also analyzed under microscopy. Representive size of the colony was photographed under high power microscopy (×100).