Extended Data Fig. 6: SPHK1 selective inhibitor mitigates established aortic muscularization in Eln(+/−) mice.
From: Sphingosine kinase 1 is integral for elastin deficiency-induced arterial hypermuscularization
(a) Transverse aortic sections from Eln(+/+) or Eln(+/−) pups at P0.5 were stained for markers of proliferation (Ki67), SMCs (SMA), and nuclei (DAPI). (b) Histograms represent aortic medial wall thickness from sections as shown in a. n = 10 mice per group. (c) Percent of SMCs that are Ki67+ from sections represented in a. n = 5 Eln(+/+) mice, n = 7 Eln(+/−) mice. (d) Experimental strategy for (e-i). Eln(+/+) or Eln(+/−) pups were injected daily with PF-543 (20 mg/kg body weight) or vehicle from P2.5 to P6.5 and analyzed at P8.5. Graphical objects were created with BioRender (https://biorender.com/). (e, h) Transverse aortic sections were stained for SMA, nuclei (DAPI) and either Ki67 in e or cleaved caspase-3 (apoptosis marker) in h. (f) Histograms represent aortic medial wall thickness from sections at P8.5 as shown in e. n = 9 Eln(+/+) mice with vehicle; n = 6 Eln(+/+) mice with PF-543; n = 8 Eln(+/−) mice with vehicle; n = 7 Eln(+/−) mice with PF-543. (g) Percent of SMCs that are Ki67+ from sections represented in e. n = 5 mice per group. (i) Percent of SMCs that are cleaved caspase-3+ from sections represented in h. n = 6 Eln(+/+) mice with vehicle; n = 5 Eln(+/+) mice with PF-543; n = 5 Eln(+/−) mice with vehicle; n = 5 Eln(+/−) mice with PF-543. Bar graphs are shown as mean ± SD. P values were determined by two-tailed unpaired Student’s t test (b, c) or one-way ANOVA with Tukey’s post hoc test (f, g, i). Scale bars, 50 μm (a, e, h).
