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. 2022 Jul 19;9:770065. doi: 10.3389/fcvm.2022.770065

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Copyright © 2022 Chakrabarti, Bhattacharya, Gebere, Johnson, Ayub, Chatzistamou, Vyavahare and Azhar.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Haploinsufficiency of Tgfb1 and Smad3 inhibits canonical and non-canonical TGFβ signaling in the aortic valve of Kl^–/– mice. (A) Western blot analysis on three different micro-dissected pooled tissues, from two to three hearts/sample, representing AV leaflets and hinge area and aortic annulus of 10–12-week-old wild-type, Kl^–^/^–, Kl^–^/^–;Tgfb1^±, and Kl^–^/^–;Smad3^± mice showing protein levels of the phosphorylated forms of canonical and non-canonical pathway molecules (pSMAD2, pSMAD3, pSMAD1/5, pp38, and pERK1/2) and their non-phosphorylated/total forms (SMAD2, SMAD3, SMAD1/5, p38, and ERK1/2). A common and independent β-actin blot (bottom) was also used for normalizing the data. (B) Densitometric quantification of phosphorylated proteins after normalization to total non-phosphorylated proteins or β-actin. Numerical data (mean ± SD) from three pooled samples are presented as scatter plots with bar. The p-values are shown on top of the histograms.