Statistical calculations were performed using GraphPad Prism version 9.5.0 (GraphPad Software Inc., San Diego, United States). Two-way ANOVA followed by Šidák`s correction for multiple comparisons was used for statistical comparisons of tissue bath data, i.e., of concentration-response curves. Statistical significance was regarded for p-values < 0.05. Data are presented as mean ± SEM.
Protein expression was analyzed semi-quantitatively by grading the expression of sGC in each tissue on a scale from 0 to 3 where 0 = no protein expression, 1 = weak protein expression, 2 = moderate protein expression and 3 = strong protein expression. The grading was performed by three blinded evaluators (MW, ED, JS) on images taken with a DS-Fi camera mounted in a Nikon 90i fluorescence microscope (Nikon Corporation, Tokyo, Japan). All images were taken after digital subtraction of background staining measured in corresponding negative controls (i.e., in which the primary antbody was excluded), using the NIS Element Imaging Software v. 4.40 (Nikon Corporation). Each evaluator graded each tissue. Subsequently, a consensus grade was determined for each tissue. Thereafter, the tissues were decoded and a non-parametric statistical calculation was carried out by running a Mann-Whitney rank test.
Protein expression was analyzed semi-quantitatively by grading the expression of sGC in each tissue on a scale from 0 to 3 where 0 = no protein expression, 1 = weak protein expression, 2 = moderate protein expression and 3 = strong protein expression. The grading was performed by three blinded evaluators (MW, ED, JS) on images taken with a DS-Fi camera mounted in a Nikon 90i fluorescence microscope (Nikon Corporation, Tokyo, Japan). All images were taken after digital subtraction of background staining measured in corresponding negative controls (i.e., in which the primary antbody was excluded), using the NIS Element Imaging Software v. 4.40 (Nikon Corporation). Each evaluator graded each tissue. Subsequently, a consensus grade was determined for each tissue. Thereafter, the tissues were decoded and a non-parametric statistical calculation was carried out by running a Mann-Whitney rank test.