Table 1.
Patient characteristics and plaque vulnerability.
Table 2.
The relative expression of miR-451a and miR-23a (ΔCt = Ct(miR-23a)-Ct(miR-451)) in v-plaque samples (v) and s-plaque samples (s).
Fig 1.
Lysis-buffer-treated versus intact plaques.
H&E staining of s- and v-plaques on shows elimination of plaque surface cells (arrows) after lysis-buffer treatment.
Fig 2.
Cells from v-plaque at passage 0 (cultured for 15 days). Phase contrast images representative of the 48 obtained PIMC cultures. Scale bars represent 40 μm.
Table 3.
The efficacy of PIMC isolation and culture from stable (n = 35) and vulnerable (n = 56) plaques.
Fig 3.
PIMC in various culturing conditions.
A. s-PIMC viability on the next day after subculturing (Day 1) and after 5-day incubation (Day 5) in various culture media or with different growth stimulators normalized to viability of s-PIMC in IMDM-FBS (first column of the corresponding panel). Mean of three replicates with standard deviation. Asterisk (*) indicates a significant difference in cell viability (P < 0.0001, T-test). B. Phase-contrast microscopy images of s-PIMC on Day 1 and Day 5. Representative images from two independent experiments are shown. Scale bars represent 20 μm.
Fig 4.
Principal component analysis of gene expression profiles.
RNA-Seq gene expression data of v-PIMC (n = 2), s-PIMC (n = 2) and cells involved in atherogenesis (vascular smooth muscle cells–HCASMC (n = 3), endotheliocytes–HUVEC (n = 3) and HAEC (n = 3), macrophages MCRPH (n = 3), chondrocytes–AchKJ (n = 2), osteoblasts HOB (n = 2) and fibroblasts–FAADV (n = 2)). PCA demonstrates the major components (PC1 and PC2) of the variance to separate PIMC from other cells.
Fig 5.
Heatmaps of cell type-specific gene expression.
RNA-Seq gene expression data of v-PIMC (n = 2), s-PIMC (n = 2) and cells involved in atherogenesis (vascular smooth muscle cells–HCASMC (n = 3), endotheliocytes–HUVEC (n = 3) and HAEC (n = 3), macrophages MCRPH (n = 3), chondrocytes–AchKJ (n = 2), osteoblasts HOB (n = 2) and fibroblasts–FAADV (n = 2)). Panels A-E show the expression of cell-type specific markers. Expression of each marker is normalized to the expression level in the corresponding specific cell type. Red: upregulation; green: downregulation.
Table 4.
Differentially expressed genes in v-PIMC (v) compared to s-PIMC (s).
Fig 6.
Immunofluorescence staining of s- and v-PIMC.
Specific proteins are stained green (AlexaFluor 488), f-actin and nuclei are stained red and blue, correspondingly. In the example column VSMC, HCC and HUVEC are stained for indicated markers. Representative images were selected from three independent experiments performed using PIMC obtained from different donors are shown. Scale bars represent 50 μm.