Fig. 5: Restoring NEXN phosphorylation corrects the pathological phenotype caused by Clk4 deficiency in vitro and in vivo. | Nature Communications

Fig. 5: Restoring NEXN phosphorylation corrects the pathological phenotype caused by Clk4 deficiency in vitro and in vivo.

From: CDC-like kinase 4 deficiency contributes to pathological cardiac hypertrophy by modulating NEXN phosphorylation

Fig. 5

a qPCR detection of Nppa (upper panel) and Nppb (lower panel) in NRVMs challenged with PE alone, Ad-Clk4 alone, PE + Ad-Clk4, or PE + Ad-Clk4 + Nexn siRNA. n = 3 biologically independent samples per group. b qPCR analysis of Nppa (upper panel) and Nppb (lower panel) expression in NRVMs transfected with Clk4 siRNA alone or in combination with Nexn-WT, Nexn-S437A, or Nexn-S437E. WT, wild-type. n = 3 biologically independent samples per group. c Representative western blots of exogenously expressed and total NEXN, as detected by Flag tag antibody and NEXN antibody respectively, in AAV NEXN-WT (AAV-WT)-, NEXN-S437E (AAV-S437E)- and NEXN-S437A (AAV-S437A)-infected hearts, AAV-GFP and wild-type mice were used as controls. d Upper panel, Schematic diagram of the AAV9 vector. Lower panel, protocol for AAV9 administration to Clk4-cKO mice. e, f Summary of echocardiographic data for the LV internal diameter, diastole (LVID, d) and LV ejection fraction (EF). n = 6 animals for GFP, n = 7 animals for WT, n = 7 animals for S437E and n = 5 animals for S437A. g Gross heart images of Clk4-cKO mice treated with AAV-WT, AAV-S437E, or AAV-S437A. Scale bar: 2.5 mm. h Heart weight-to-body weight ratios (HW/BW). n = 5 animals per group. i Heart weight-to-tibial length ratios (HW/TL). Data are presented as the means ± S.E.M. n = 5 animals per group. For a, b, e, f, h, and i, statistical analyses were performed using one-way ANOVA and Dunnett multiple comparisons test. Data are presented as the means ± S.E.M.; P values are shown in each graph. Source data are provided as a Source Data file.

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