Figure 4.

Plastic cages/water bottles, without the addition of BPA in water, increase mast cell numbers and pericardial degranulation in BALB/c females. Female BALB/c mice were housed in glass or plastic cages with glass or plastic water bottles and no soy in food or bedding for 3 weeks prior to ip infection with CVB3 to induce myocarditis. The drinking water did not contain BPA. Mice were injected ip with 10³ PFU of CVB3 ip on day 0 and hearts were harvested at day 10 pi during acute myocarditis. (A) Relative gene expression (RGE) of mast cells (cKit/ CD117) vs. the housekeeping gene Hprt was analyzed in whole hearts by qRT-PCR at day 10 pi comparing mice in glass vs. plastic cages (glass n = 10, plastic n = 10). (B–F) Toluidine blue was used to detect mast cell granules by histology and the total numbers of MCs in the heart section were normalized to the size of the heart using an eyepiece grid [(50); glass n = 10, plastic n = 10]. (A,B) Data show the mean ± SEM using the Mann-Whitney rank test of total mast cells (NOT and Degran). (C–F) Two-way ANOVA found significant results for degranulation (p < 0.0001) and BPA (p = 0.0006). P-values depict ad-hoc analyses comparing glass to plastic by Tukey's multiple comparison.