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Fig. 4 | Respiratory Research

Fig. 4

From: Functional changes in long-term incubated rat precision-cut lung slices

Fig. 4

Endothelin-1 (ET-1)-induced bronchoconstriction in long-term cultivated rat precision-cut lung slices (PCLS). PCLS with intact airways and surrounding epithelium were used in measurements of ET-1-induced bronchoconstriction on measuring days. Different concentrations of ET-1 (10–9–10–6 M) were added to the slice, constriction was measured via microscope. Images were analysed with ImageJ. For each timepoint, measurements were performed in duplicates, n = 4, the same slices were used for measurements with ET-1 on every measuring day. Data is expressed as mean ± standard deviation (SD). First image of non-constricted airway of each measuring day was set as 100%. ET-1-induced reduction of airway area was expressed as percentage of the initial airway area (%-IAA) (A). Using GraphPad Prism 9, nonlinear regression with a four-parameter logistic equation was conducted from data of ET-1-induced bronchoconstriction (10–9–10–6 M). Maximal constriction (= efficacy) (B), maximal airway area (C) and half-maximal response (EC50) (D) were calculated. Analysis was performed based on control image of every measuring day ( ) and that of the first measuring day (). Statistics were carried out using the general mixed model analysis (PROC GLIMMIX, SAS 9.4, SAS Institute Inc., Cary, NC, USA), statistical differences to day 1 are indicated by asterisks (**0.001 ≤ p < 0.01, ***p < 0.001)

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