Fig. 7: SOX9 drives metastatic-specific cell dormancy through RA-response. | Nature Communications

Fig. 7: SOX9 drives metastatic-specific cell dormancy through RA-response.

From: Oncogenic enhancers prime quiescent metastatic cells to escape NK immune surveillance by eliciting transcriptional memory

Fig. 7: SOX9 drives metastatic-specific cell dormancy through RA-response.The alternative text for this image may have been generated using AI.

a Representative images and quantification of p27 positive cells in MD expressing shGFP (Ctrl) or shSOX9 (KD SOX9) in steady state or after 72 h treatment with atRA. p27, white; DAPI, blue; scale bar = 20 µm. b Representative histograms of FACS analysis of Ctrl MD and KD SOX9 MD cells showing CellVue Maroon staining intensity (APC-A) 8 days (T1) or just after staining (T0), in the indicated conditions; barplot of the percentage of quiescent cell is shown. c Schematic representation of the experimental setup for NK cytotoxicity assays. (created with BioRender.com). d Average percentage of NK-mediated cytotoxicity of cycling and quiescent MD cells expressing either shGFP (Ctrl) or shSOX9 (KD SOX9), for different effector:target (E:T) ratios. e Barplots of relative gene expression levels of NK activating (green) and inhibitory (red) ligands in bulk and quiescent (p27 + ) MD cells. f Average percentage of NK-mediated cytotoxicity of quiescent MD cells for different effector:target (E:T) ratios, after vehicle or Navitoclax treatments. g Schematic representation of the experimental setup for metastatic colonization assay (created with BioRender.com). h DTCs and metastatic foci distribution within the lungs of nude mice injected with Ctrl or KD SOX9 MD cells. Top, tile scans of entire lung sections; scale bar: 1 mm. Bottom, representative images of metastatic foci (corresponding to i and ii from top image; scale bar = 1 mm) and quiescent DTCs (corresponding to iii-v from top image; scale bar = 50 µm; further zoom scale bar = 10 µm). mCherry: yellow; p27_Venus: green; DAPI: blue. i Barplots of scattered DTCs and metastatic foci in lungs of mice injected with Ctrl or KD SOX9, normalized to the lung area analyzed (n = 16; data combined from two experimental groups). j Barplots of the percentages of cycling or quiescent cells retrieved as scattered DTCs or lung metastases in nude mice (n = 16; data combined from two experimental groups). k Schematic representation of the experimental setup for metastatic colonization assay in humanized mouse models. (created with BioRender.com) l DTCs and metastatic foci distribution within the lungs of NSG mice injected with Ctrl or KD SOX9 MD cells and with or without NK-92 cells. Right, tile scans of entire lung sections; scale bar = 1 mm. Left, representative images of metastatic foci (corresponding to i and iii from top image; scale bar = 1 mm) and quiescent DTCs (corresponding to ii, iv, v, and v from top image; scale bar = 50 µm). Human mitochondria: yellow; p27_Venus: green; DAPI: blue. m Barplot of scattered DTCs and metastatic foci normalized to the lung lobe area analyzed in lungs of NSG mice injected with Ctrl or KD SOX9 MD cells, and with or without NK-92 cells, (n = 15; data combine from three experimental groups). n Barplots of the percentages of cycling or quiescent cells retrieved as scattered DTCs and metastatic foci in the lungs of NGS mice (n = 15; data combined from three independent experimental groups). The barplots in (a, b, e) are means of three independent biological replicates ± S.E.M. The mean values ± S.E.M in (d) and (f) were retrieved from three independent biological replicates. Statistical significance was determined by one-tailed unpaired student’s t-test.

Back to article page