Fig. 3

A Representative picture of the silver stained SDS polyacrylamide gel. Lanes represent the saliva samples mixed with the following substances: a. HD 4-times diluted, b. HD 5-times diluted, c. HD 6.67-times diluted, d. HD 10-times diluted, e. HD 20-times diluted, f. PBS, g. 10% ethanol, h. native saliva, M: low molecular weight protein marker. Yellow rectangles show the area in each lane the density was measured using Fiji (ImajeJ2) software. Numbered red rectangles show the bands selected for pixel intensity measurements in each lane to compare effects of different treatments. B Calibration of molecular weight based on the relative migration distance of the low molecular weight markers measured in pixels on the gel photograph. Black points show the molecular weight markers, whereas red points show the molecular weight (log₁₀) of the selected bands based on their migration distance. y = -0.001055x + 2.051 is the equation of the linear regression, R² = 0.9786 represents the goodness of fit, p = 0.0002. C Integrated pixel density of the separate lanes on the SDS-PAGE (yellow rectangle in A). Bars represent the mean ± SEM of two technical replicates (two different gels loaded with the same samples). Ordinary one-way ANOVA with Dunnett’s multiple comparisons test. D Integrated pixel density of the selected bands (1 to 7, red rectangles in A) on the SDS-PAGE from each lane. Two-way ANOVA with Dunnett’s post hoc test, comparing every treatment to the native saliva. Bars represent the mean ± SEM of two technical replicates (two different gels loaded with the same samples)