Figure 3.

Cortical mRNA E/I analysis and Satb2 translational inhibition

(A) EISA of CITF mRNAs. Outline shows different ratios of exonic and intronic sequences in relation to mRNA stability as rationale at the basis of EISA. Box plots show the ratio of E/I read counts, and intron read counts, for distinct CITFs and Actb (constitutive control gene) in cortical progenitors at different in vivo embryonic times.

(B) Density plot of E/I ratio fold change between E13.5 and E15.5.

(C) qRT-PCR evaluation of Argonaute (AGO)-interacting Satb2 mRNA. Values on y axis report the ratio of RT-PCR-detected, immunoprecipitated Satb2 mRNA with respect to the input (AGO RIP). GFP RIP, control immunoprecipitation with anti-GFP Ab. N = 3 independent experiments. Asterisk indicates p-value = 0.049 (Student’s t test).

(D) Expression of Satb2 3′ UTR-bearing GFP reporter after lipofection in corticalized mESCs. N = 3 independent experiments. Cells were transfected 48 h before the time of analysis indicated in labels. Asterisk indicates p-value = 0.000061 (Wilcoxon signed rank test).