Figure 4. Actin remodeling and adhesion... | American Society of Hematology

$Actin remodeling and adhesion defects in NK cells from patients with MM. (A) Heatmap of leading-edge genes standardized expression from the “regulation_of_actin_cytoskeleton” KEGG pathway across NK clusters or disease state (see GSEA analysis in Figure 2G-H). (B) FACS plots and graph showing conjugates between cell trace violet-labeled thawed HD or NDMM NK cells and RPMI8226 stained with PKH67 and coated or not with isatuximab at a ratio of 2:1 for 15 minutes. (C) FACS plots and graph showing PKH67 acquisition (trogocytosis) by thawed HD or MM NK cells in the presence of RPMI8226, stained with PKH67 and coated or not with isatuximab at a ratio of 2:1 for 4 hours. (D) FACS plots and graphs showing total or open LFA-1 expression by thawed HD and NDMM NK cells on ICAM-1 coated wells for 4 hours. (E) Representative pictures showing actin ring formation (phalloidin; green) and open LFA-1 (m24; red) by thawed MM or HD NK cells plated on ICAM-1–coated wells for 20 minutes. (F) Graph showing the actin ring area, length/width ratio and LFA-1 spots per rings in the indicated conditions. Each dot represents the mean of at least 50 NK cells from each donor. (G) Phalloidin expression by thawed PB Lin–CD56+ NK cells from HD, NDMM, and relapsed/refractory MM analyzed by flow cytometry. (H) Phalloidin expression by the indicated NK cells subsets from NDMM identified as in Figure 3B. Each dot represents an independent donor. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; Mann-Whitney test or ANOVA with Tukey post hoc test analysis.$

Figure 4.

Actin remodeling and adhesion defects in NK cells from patients with MM. (A) Heatmap of leading-edge genes standardized expression from the “regulation_of_actin_cytoskeleton” KEGG pathway across NK clusters or disease state (see GSEA analysis in Figure 2G-H). (B) FACS plots and graph showing conjugates between cell trace violet-labeled thawed HD or NDMM NK cells and RPMI8226 stained with PKH67 and coated or not with isatuximab at a ratio of 2:1 for 15 minutes. (C) FACS plots and graph showing PKH67 acquisition (trogocytosis) by thawed HD or MM NK cells in the presence of RPMI8226, stained with PKH67 and coated or not with isatuximab at a ratio of 2:1 for 4 hours. (D) FACS plots and graphs showing total or open LFA-1 expression by thawed HD and NDMM NK cells on ICAM-1 coated wells for 4 hours. (E) Representative pictures showing actin ring formation (phalloidin; green) and open LFA-1 (m24; red) by thawed MM or HD NK cells plated on ICAM-1–coated wells for 20 minutes. (F) Graph showing the actin ring area, length/width ratio and LFA-1 spots per rings in the indicated conditions. Each dot represents the mean of at least 50 NK cells from each donor. (G) Phalloidin expression by thawed PB Lin^–CD56⁺ NK cells from HD, NDMM, and relapsed/refractory MM analyzed by flow cytometry. (H) Phalloidin expression by the indicated NK cells subsets from NDMM identified as in Figure 3B. Each dot represents an independent donor. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; Mann-Whitney test or ANOVA with Tukey post hoc test analysis.

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