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. 2023 Jun 8;24(12):9909. doi: 10.3390/ijms24129909

Figure 4.

Figure 4

Software-based analyses of CD163/CD8 EF ratio. (a) Sequential IHC pseudo-color images of CD163 (cyano) and CD8 (red) cells within EF areas. Scale bars 200 μm. (b) Representative bright-field IHC CD21 ROI image used to create black and white masks for EF area segmentation and the segmentation results are indicated as a scatter dot plot of the percentage of EF occupied area across 5 ROIs for the four selected patients (PD_N: #ID8, #ID20; PD_Y: #ID30, #ID14). Statistical analysis was performed using 2-way ANOVA with a Šídák’s multiple comparisons test. (c) For each patient, an image cytometry-based cell population analysis for the lymphoid (CD8) and myeloid (CD163) biomarkers was obtained using the 5 ROIs previously described. Percentage of the CD8+ and CD163+ population are highlighted in red. (d) Violin Plot of percentage of CD163/CD8 EF ratio between 10 PD_N ROIs and 10 PD_Y ROIs. PD, progressive/recurrent disease; ROIs, regions of interest.