Figure 2
From: Preserving Airway Smooth Muscle Contraction in Precision-Cut Lung Slices
The DMF12-Ins medium has no effect on the viability of PCLS but preserves smooth muscle myosin heavy chain (SMMHC) expression in ASM. (a) Representative confocal images of live-dead stained PCLS (left panel) and a summary plot of living to dead cell ratio (right panel) in PCLS from the conventional DMF12 group and the new DMF12-Ins group at days 1 and 15. Dead cells are labeled with red fluorescence. Scale bar = 50 μm. Each column represents the mean ± SEM of 6–7 slices from 3 mice. (b) Representative flow cytometry analyses and summary plot of viable cell percentage at days 1 and 15 in DMF12 and DMF12-Ins groups. Each measurement represents the mean ± SEM of 4 independent experiments. (c) Colorimetric cell metabolic analyses of PCLS at days 1, 7, and 12 in two culture conditions. Each dot represents mean ± SEM of 14 slices from 4 mice. *p < 0.05 by Sidak’s test following two-way ANOVA. (d) Representative images (upper panel) of the PCLS prepared from SMA-GFP transgenic mice and the summary plot of airway SMA fluorescent intensity (lower panel) on days 1 and 15 in the DMF12 and DMF12-Ins groups. (e) Representative SMMHC immunofluorescent images (upper panel) and the summary plot of fluorescence intensity (lower panel) on days 1 and 15 in the DMF12 and DMF12-Ins groups. The expression level of SMA and SMMHC was presented as a ratio to the average level of the respective protein in the DMF12 PCLS culture on day 1. The arrow marks the airway. The bar graph represents mean ± SEM of 7–8 slices from 4 mice. *p < 0.05, by Tukey’s test following 2-way ANOVA.
