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. 2019 Jan 4;20(1):168. doi: 10.3390/ijms20010168

Figure 4.

Figure 4

Effects of PACAP and/or oxidative stress and/or mechanical stress (MS) on metachromatic cartilage formation. Metachromatic cartilage areas in six-day-old cultures were visualized with DMMB dissolved in 3% acetic acid. Metachromatic (purple) structures represent cartilaginous nodules formed by many cells and a cartilage matrix rich in polyanionic GAG chains. Original magnification was 4×. Scale bar, 500 µm. Optical density (OD625) was determined in samples containing toluidine blue (TB) extracted with 8% HCl dissolved in absolute ethanol. Absorbance and means of three independent experiments (±standard error of the mean) are shown in the figures. Asterisks indicate significant differences compared with the six-day cultures (p < 0.05, one-way ANOVA followed by Tukey’s HSD test). * shows significant differences between Control cultures and treated samples; ** shows significant differences between single-H2O2 treatment and combined treatments.