Fig. 4: ADM is transcriptionally regulated by mechano-YAP/TAZ axis and independent of PDE5i.
From: The role of mechano-regulated YAP/TAZ in erectile dysfunction
a Volcano plot showing the altered genes in primary penile SMCs treated with siYAP/TAZ vs siCo. b The expression of Adm in primary penile SMCs treated with siYAP/TAZ. Data are the mean ± sem of n = 4 biologically independent samples. c Western blots were used to assess the expression of YAP, TAZ and ADM in primary penile SMCs derived from Yapfl/fl;/Tazfl/fl mice treated with Ad-Cre to induce YAP/TAZ knockout, the experiments were repeated three independent times with similar results. d ChIP-PCR assesses the interaction between YAP and the ADM promotor in SMCs, n = 2 biologically independent samples. e Immunohistochemistry assessing the expression of Adm from wild-type and Y/T cKO mice, the experiments were repeated three independent times with similar results. f, g qRT-PCR and Immunohistochemistry assessed the expression of Adm from sham rats (n = 12) and 5 days (n = 7), 14 days (n = 8), 28 days (n = 6) and 60 days (n = 8) indicated time points of the BCNI-induced rat ED models. h, i qRT-PCR and Immunohistochemistry assessed the expression of Adm from control (n = 5) and 1 month (n = 4), 2 months (n = 5), 3 months (n = 4) and 5 months (n = 5) indicated time points of the IR-induced rat ED models. j, k Immunofluorescence of ADM (green) and DAPI (blue) in SMCs replated on a high stretch (n = 48) or low stretch (n = 42) ECM, each point represents an area examined over 3 independent experiments. l qRT-PCR assessing the expression of Adm in SMCs treated with the indicated mechanotransduction inhibitors (n = 3). m qRT-PCR assessed the expression of Adm in SMCs treated with PDE5i or stretch when plated in dense states (n = 4). Immunofluorescence (o) and quantification (n) of ADM (green) in SMCs treated with PDE5i in medium (PDE5i untreated, n = 45; PDE5i treated, n = 46) or high (PDE5i untreated, n = 51; PDE5i treated, n = 53) stretch ECM, each point represents an area examined over 3 independent experiments. p ChIP-PCR assesses the interaction between YAP and the ADM promotor in SMCs treated with the latrunculin A (Lat.A) or PDE5i on the basis of high stretch ECM (n = 2 biologically independent samples). q Expression of Adm in SMCs treated with Ble or Jas (n = 4). r Expression of Adm in SMCs treated with Lat.A or Myo A (n = 3). Dot plots indicate mean (middle line) and all points. Bar charts are presented as the mean ± sem. The statistical analysis was calculated by two-sides unpaired Student’s t test, the confidence interval is 95%. Each experiment was repeated three independent times with similar results. The point represents a rat or detected cell area over 3 biologically independent experiments with similar results. Scale bars, 20 µm. Source data are provided as a Source Data file.
