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Figure 4 | BMC Cancer

Figure 4

From: Regulation of p14ARF expression by miR-24: a potential mechanism compromising the p53 response during retinoblastoma development

Figure 4

Role of miR-24 in retinoblastoma ARF regulation. (A) Real-time quantitative PCR analysis of miR-24 expression in human fetal retina (FR) and adult retina (HR 50 and 108), RB1 -/- retinoblastoma tumors (RB 2133, 2345, 2362, 3103), RB1 -/- retinoblastoma cell lines (RB 247, 381, 383, 1021, WERI-Rb1 (WERI), Y79), RB1 +/+ retinoblastoma cell lines (RB 3823, 522) and other cell lines (HeLa, SKOV3, HEK-T, SaOS2, OVCAR3). N = 3. (B) Comparison of normalized ARF protein expression (values taken from ARF protein/ARF mRNA expression, as depicted in Figure 1C) to miR-24 expression in human fetal retina (FR), human adult retina (HR 50), RB1 -/- retinoblastoma cell lines (RB 247, 381, 1021, WERI-Rb1 (WERI), Y79), RB1 +/+ retinoblastoma cell lines (RB 3823, 522) and other cell lines (HeLa, SKOV3, HEK-T, SaOS2, OVCAR3). (C) Ratio of miR-24 expression (normalized to RNU44), relative to normalized ARF protein expression (values taken from ARF protein/ARF mRNA expression, as depicted in Figure 1C). (D) Quantitative real-time PCR analysis of miR-24 after WERI-Rb1 cells were transiently transfected with anti-miR-24 siRNA or control siRNA for 72 hours, and presented as fold change normalized to RNU44 expression, relative to control siRNA; N = 3, t-test *P < 0.05. (E) Quantitative real-time PCR analysis of ARF mRNA after anti-miR-24 siRNA or control siRNA transfection, N = 3. (F) ARF expression measured by immunoblot in response to control siRNA and anti-miR-24 siRNA transient transfection of WERI-Rb1; ARF protein levels in HeLa cells are shown for positive ARF protein expression. (G) Graph represents fold change in ARF protein in WERI-Rb1 cells normalized to β-tubulin, relative to control siRNA. N = 3, t-test *P < 0.05.

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