“The final staining extent score was determined by multiplying the percentages of collagen I-positive area with the intensity score.. Additionally, we determined the number of CD68+ clusters by using the CD68 antibody (OriGene, MA, USA; UM800047, 1:200 dilution) to evaluate the degree of macrophage infiltration.. A CD68+ cluster was defined as more than 60 CD68+ cells per field at 200× magnification.”
“Sections (thickness, 3 µm) from the carotid arteries of the mice that were sacrificed on days 3, 7, 14 and 28 following ligation were obtained at 1 mm proximal to the ligature.. The common carotid arteries were cut longitudinally and antibodies against cluster of differentiation (CD)45 monoclonal antibody (mAb; 1:50; cat. no. ZM-0213; ZSGB-BIO), CD68 mAb (1:100; cat. no. UM800047; OriGene Technologies, Inc., Rockville, MD, USA), proliferating cellular nuclear antigen (PCNA) mAb (1:50; cat. no. TA301137; Cell Signaling Technology, Inc., Danvers, MA, USA) and smooth muscle cell (SMC) α-actin mAb (1:100; OriGene Technologies, Inc.) were used.. Antigen retrieval was performed for cross sections with citric acid monohydrate (BD Pharmingen) at 95°C for 20 min.”
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