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. 2023 Jul 18;14:4312. doi: 10.1038/s41467-023-39965-6

Fig. 5. Enhancing RBM20-TNPO3 interaction restores nuclear localization and splicing in vitro and in vivo.

Fig. 5

a Representative images of RBM20 localization upon overexpression (OE) of eGFP-TNPO3 in iPSC-CMs. Red arrows point at cells transduced with eGFP-TNPO3, n = 3. b DAPI:RBM20 colocalization based on the data shown in (a). Each dot represents a Pearson coefficient for at least five cells, n = 3. TNPO3 overexpression effect’s P value < 0.0001. c qPCR analysis of TTN exon 242 splicing (TNPO3 overexpression effect’s P value = 0.057); and d IMMT exon 6 splicing (TNPO3 overexpression effect’s P value = 0.015) in iPSC-CMs. Isoform expression was normalized to GAPDH and displayed as fold change versus the WT line (first replicate) without TNPO3 OE (means with standard errors, two biological replicates with two technical replicates for each). b–d TNPO3’s overexpression effect’s P values were calculated with Two-way ANOVA with two-tailored Tukey’s HSD post test; comparison of WT vs P633L + TNPO3 - two-tailored t-test. e Scheme of Tnpo3 OE in vivo. f qPCR analysis of Tnpo3 expression in n = 4 WT mice, n = 4 P635L+/+ mice injected with PBS, or n = 4 P635L+/+ mice injected with Tnpo3. Data is normalized to Gapdh expression and displayed as fold change versus one of the WT mice with standard errors. g Representative images of RBM20 localization in WT mice, P635L+/+ mice injected with PBS, or P635L+/+ mice injected with Tnpo3. Red arrows point at characteristic nuclear foci formed by RBM20, n = 4 mice. h qPCR analysis of Ttn splicing in n = 4 WT mice, n = 4 P635L+/+ mice injected with PBS, or n = 4 P635L+/+ mice injected with Tnpo3. Data is normalized to Gapdh expression and displayed as fold change versus one of the WT mice with standard errors. i RT-PCR of Ttn splicing and Gapdh expression in n = 3 WT mice, n = 3 P635L+/+ mice injected with PBS, and n = 3 P635L+/+ mice injected with Tnpo3. Red arrows point at Ttn isoforms expressed in WT mice. Each boxplot of this figure (b, f, h) displays quartiles Q1, Q2 (center), and Q3, with whiskers extending to the furthest data point within 1.5 times the IQR. Actual P values are shown in the source data file.