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Fig. 2 | BMC Medical Genetics

Fig. 2

From: The role of p19 and p21 H-Ras proteins and mutants in miRNA expression in cancer and a Costello syndrome cell model

Fig. 2

Gene expression and alternative splicing regulation by H-Ras proteins. a Scheme of the Taqman assays used to determine endogenous p19, p21, and endogenous total H-Ras mRNA expression. E3-IDX, E3-E4A, and E4A-E4B recognize endogenous p19 H-Ras, p21 H-Ras, and both p19 and p21 H-Ras, respectively. Taqman assays were performed with E3-E4A and E4A-E4B for overexpressed p19 and with E3-IDX and E4A-E4B for overexpressed p21mut. b Endogenous total H-Ras (Taqman E4A-E4B) and p21 (Taqman E3-E4A) mRNA levels change upon overexpression of p19 and p19mut. The results are presented as fold changes with respect to basal levels obtained for empty vector transfections, which were set to 1 (no change). Graphic: the percentage of alternative splicing p21/endogenous total H-Ras (Taqman E3-E4A/Taqman E4A-E4B) was obtained for each individual experiment and standard deviations of three separate experiments calculated. (−) is the % p21 in transfections with empty vector set to 33 %; () p19 and () p19mut, % is the p21 in HeLa cells overexpressing p19 and p19mut, respectively. () p19, P = 0.07; () p19mut, P = 0.08. c Endogenous total H-Ras (Taqman E4A-E4B) and p19 (Taqman E3-IDX) mRNA level changes upon overexpression of p21Q61L. The results are presented as fold changes with respect to the basal levels obtained for empty vector transfections, which were set to 1 (no change). Graphic: the percentage of alternative splicing p19/total H-Ras (Taqman E3-IDX/Taqman E4A-E4B) was obtained for each individual experiment and standard deviations of three separate experiments calculated. (−) is the % of p19 in transfections with empty vector was set to 15 %; ()p21mut is the % of p19 after expression of the p21mut (p21Q61L). P = 0.09. Hela cells line were used in all the experiments stated above

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