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The percentage of cells with 2C, 4C and intermediate levels of DNA content was calculated using the WinMDI program.
DNA content was calculated as mean ploidy by multiplying the nuclear ploidy level by the number of nuclei in each ploidy class.
The DNA content was calculated using a standard curve.
The percentage of hypodiploid cells (<2C DNA content) was calculated from the total number of cells.
DNA content was calculated using the formula (G1 cells + 2 × G2 cells)/(G1 cells + G2 cells) [ 13 ].
In addition, total protein content per DNA content was calculated, since DNA content is commonly used for cell number normalization.
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The protein level of osteolytic factors normalized by the DNA contents were calculated as: [(concentration of erlotinib-treated wells − mean concentration of cell-free wells)/DNA contents of erlotinib-treated wells]/[(concentration of vehicle-treated wells − mean concentration of cell-free wells)/DNA contents of vehicle-treated wells] × 100.
G+C content was calculated from the proteome as DNA.
RPG4/GC or PG4 DNA density normalized for GC content was obtained by dividing the PG4 DNA density by GC content of a given region; GC content was calculated using in-house software.
DNA content in the pellets was analyzed with a Quant iT ds Pico Green DNA Assay Kit (Invitrogen, Karlsruhe, Germany) according to manufacturers' protocols, and GAG/DNA content was calculated.
The relative water content was calculated.
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