Fig. 8

Gene expression of rat native lung tissue compared to rat precision-cut lung slices (PCLS). Native lung tissue (Lobus accessorius) and PCLS of the preparation day were analysed for messenger ribonucleic acid (mRNA) expression of endothelin-1 (Edn1, A), endothelin-2 (Edn2, B), endothelin-3 (Edn3, C), endothelin receptor type A (Ednra, D), endothelin receptor type B (Ednrb, E), thromboxane A₂ receptor (Tbxa2r, F), thromboxane A synthase 1 (Tbxas1, G), acetylcholinesterase (Ache, H), muscarinic acetylcholine receptor M1 (Chrm1, I), muscarinic acetylcholine receptor M2 (Chrm2, J) and muscarinic acetylcholine receptor M3 (Chrm3, K) in relation to a reference gene index (Ref. gene index) consisting of ribosomal protein lateral stalk subunit P0 (Rplp0) and beta-actin (Actb). For this purpose, native lung tissue and PCLS of the preparation day were snap frozen in liquid nitrogen after withdrawal and PCLS preparation, respectively. Ribonucleic acid (RNA) isolation was performed by a combination of phenol–chloroform extraction and magnetic-bead technique. Afterwards, reverse transcription (RT) and quantitative polymerase chain reaction (qPCR) were performed. Measurements were performed in triplets, an average of 6–8 slices (n = 5) and native lung tissue of Lobus accessorius (n = 9) were snap frozen in liquid nitrogen, of which 100 mg of the slices and 20 mg of the lobes, respectively, were used for RNA isolation. RT-qPCR was performed with an input of 230 ng RNA. Data is expressed as mean + standard deviation (SD), dots represent individual data points. Statistics were carried out using the general mixed model analysis (PROC GLIMMIX, SAS 9.4, SAS Institute Inc., Cary, NC, USA), statistical differences to the control group (Lobus accessorius) are indicated by asterisks (*0.01 ≤ p < 0.05; **0.001 ≤ p < 0.01, ***p < 0.001, ns no significance)