Figure 4.

Role of PDE5 in regulating the accumulation of ST-stimulated cGMP in HEMs. (a) PDE5 KD efficiency screening by quantitative RT-PCR in three different shRNA clones. Clone 330 was used in all subsequent experiments. (b) cGMP quantitation in apical, intracellular, and basolateral pools from ST-exposed HEMs (1 nM ST, 1 h). IBMX was not present in these experiments. Scr, scrambled shRNA enteroid line; PDE5 KD, PDE5 shRNA-expressing enteroid line. Vardenafil (10 μM) was included in designated samples. Data points represent technical replicates from three independent experiments and graph bars are mean ± SEM. Statistical significance was determined by ANOVA. *p < .05; ***p < .001.