Figure 3.

Comparison of Atheroma Macrophage and Smooth Muscle Cell Presence, and Plaque Cathepsin RNA Expression in DCB-PTA, PTA, and Sham-PTA Subjects, as Well as in Vitro Assessment of the Effects of Paclitaxel on Cathepsin Expression in Human Aortic Smooth Muscle Cells

Lesion sections from representative DCB-PTA, PTA, and sham-PTA animals underwent (A and B) RAM-11 macrophage (n = 143 sections) and (C and D) alpha-actin smooth muscle cell (n = 79 sections) immunohistochemical detection. The percent positively stained area in lesions was assessed; balloon dots are actual values per section; bars demonstrate median (25th and 75th percentiles). (E) Gene expression normalized to the expression of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (housekeeping gene), displayed using comparative Ct method (2^-ΔΔCt). mRNA expression of cathepsin genes measured by quantitative polymerase chain reaction. Verification of expression demonstrated by immunoblotting of cathepsin B, after protein extraction. Statistical comparisons made using Kruskal-Wallis H test, with a post hoc analysis using Dunn’s test of multiple comparisons. (F) In vitro human aortic smooth muscle cells were incubated with graded concentrations of paclitaxel, and then underwent RNA extraction to assess cathepsin transcripts. n = 4, bars demonstrate median (25th and 75th percentiles). ∗∗∗p < 0.001. Abbreviations as in Figure 1.