Chlamydia RNA in the aorta tissues. (a) Experimental design for Cpn infection of ApoB100only/LDLR−/− mice and 16SrRNA transcript detection in pooled aorta samples (two pools of 3 aortas from 6 mice) of one time (1x Cpn) and 3 times (3x Cpn) Cpn-infected and noninfected mice (“+”: aorta samples tested positive; “−”: aorta samples tested negative for expression of 16SrRNA by qRT-PCR). ((b)A) Identification of the RT-PCR-amplified Cpn 16SrRNA cDNA in the aorta of Cpn-infected ApoB100only/LDLR−/− mice. Melt curves show Cpn 16SrRNA amplicons in (1) Cpn-infected McCoy cells (positive control); (2), (3) in aorta samples tested 1 week after single Cpn infection; (4) in aorta samples 5 weeks after third Cpn infection, (5), (6) in aorta samples 4 weeks after a single Cpn infection; ((b)B) β-actin amplicons in all tested aorta samples and Cpn-infected McCoy cells. (c) Expression of Cpn 16SrRNA normalized to the expression level of mouse β-actin at 1 week after single Cpn infection was set as 1 and relative mouse β-actin-normalized Cpn 16SrRNA expression level in all pooled aorta samples giving positive 16SrRNA signal was calculated. The measurements were repeated two times with the same results.