Fig. 5: Multiple sites for nELAVL proteins are present on the mRNAs encoding the calcium channel subunits.
From: CYFIP1 governs the development of cortical axons by modulating calcium availability
a Histograms indicating the frequency of nELAVL binding sites (represented as normalized Peak Height) on the coding sequence and 3’ untranslated region (UTR) of human hCACNA1C, hCACNA1E and hCACNA1I mRNAs, as found in previous CLIP-seq published data66. b Histograms representing the frequency of predicted binding sites of HuD and HuR on the 5’ UTR, coding sequence and 3’UTR of mouse mCacna1c, mCacna1e and mCacna1i mRNAs, generated using RBPmap67. c HuD/Elavl4 RNA immunoprecipitation (RNA-IP) from DIV 3 WT and Cyfip1+/- cortical neurons. Histograms represent calcium channel mRNAs enrichment, calculated as the ratio over the positive control (mDlg4) (WT n = 4 embryos, Cyfip1+/- n = 4 embryos; mean ± SEM, Two-tailed Multiple Mann-Whitney test; mCacna1c mRNA p = 0.0285, mCacna1e mRNA p = 0.0285, mCacna1i mRNA p = 0.8857). d HuR/Elavl1 RNA-IP from DIV 3 WT and Cyfip1+/- cortical neurons. Histograms represent calcium channel mRNAs enrichment, calculated as the ratio over the positive control (mDlg4) (mean ± SEM, Two-tailed Multiple Mann-Whitney test; mCacna1c mRNA p = 0.0310, mCacna1e mRNA p = 0.2270, mCacna1i mRNA p = 0.6289). e Histograms representing the mRNA decay after transcriptional shutdown with Actinomycin D in WT and Cyfip1+/- DIV 3 cortical neurons. mRNA expression levels of each calcium channel subunit mCacna1c, mCacna1e and mCacna1i were normalized to mH3f3 mRNA at each time point (0, 2, and 4 h after Actinomycin D treatment (mCacna1c: WT n = 3 embryos, Cyfip1+/- n = 4 embryos; Two-way ANOVA, F(2, 15) = 6.834, p = 0.0078; time p < 0.0001, genotype p = 0.0137, interaction p = 0.0078; Sidak’s multiple comparisons test, 2 h p = 0.9957, 4 h p = 0.0010; mCacna1e: WT n = 3 embryos, Cyfip1+/- n = 4 embryos; Two-way ANOVA, F(2, 15) = 6.697, p = 0.0083; time p = 0.0002, genotype p = 0.0006, interaction p = 0.0083; Sidak’s multiple comparisons test, 2 h p = 0.1052, 4 h p = 0.0003; mCacna1i: WT n = 3 embryos, Cyfip1+/- n = 4 embryos; mean ± SEM, Two-way ANOVA, F(2, 15) = 1.905, p = 0.1831; time p < 0.0001, genotype p = 0.0218, interaction p = 0.1831; Sidak’s multiple comparisons test, 2 h p = 0.2958, 4 h p = 0.0454). Source data are provided as a Source Data file.
