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. 2019 Jul 25;4(14):e124716. doi: 10.1172/jci.insight.124716

Figure 7. RAS SnCs evade immune clearance with high MMP expression and robust NKG2D ligand shedding.

Figure 7

(A) Survival of RAS-induced senescent WI-38, IMR-90, and HCA-2 cells with active or inactive p53 (p53–) assessed by cell number after 10 days of direct coculture with PBMCs. (B) mRNA levels of NKG2D ligands and MMPs in RAS SnCs were measured by quantitative real-time PCR, and normalized to the average expression in control and p53-deficient presenescent cells. The range of the color scale is 64-fold greater than in Figure 2, A, C, E, and G, and Figure 5A. The ratio of the average expression levels of MMPs relative to NKG2D ligands is ~130 in RAS SnCs compared with ~7 in XRA or REP SnCs. (C) MMP-1, -3, -10, -12 immunofluorescence comparing PRE, XRA, and RAS-induced senescent WI-38 cells. Original magnification, ×20. (D) Detection of soluble MICA by ELISA in conditioned media of RAS-induced senescent WI-38, IMR-90, and HCA2 cells (p53-deficient or WT). Left: MICA levels in monocultures (naive), with SEN (XRA) and PRE cells for comparison. Right, MICA levels in supernatants collected after 10 days of direct coculture with PBMCs (persistent SnCs). In A and D, box plot length: 25% and 75% of data; centerline: median; whiskers: 25% – (or 75% +) 1.5 × IQR; dots: outliers; color bars: average in A, median in D. (E) Proportion of cells with different MMP3 levels quantified from immunofluorescence of naive and persistent RAS SnCs (top: p53 WT; bottom: p53-deficient; quantified as in Figure 5D).