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Conditioned-medium (PITX2-CM) is collected after transient transfection with PITXA, -B, or -C. Freshly plated SKOV-3 cells are incubated for 8 h with PITX2-CM alone or in combination with 30 ng/ml recombinant DKK1 followed by isolation of RNA. Q-PCR assay is performed from RNA with the primers of CCND1 (a), c-MYC (b), and PCNA (c) genes. The comparative expression is indicated as change in fold in the y axis (mean ± S.E.). * represents p < 0.05.
