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. 2020 Sep 16;11:4660. doi: 10.1038/s41467-020-18189-y

Fig. 7. Soluble CD109 is a mediator of HDAC1-derived intercellular signals from core to edge GBM cells.

Fig. 7

a In vitro cell growth assay of edge-like 157 GBM spheres treated with CM from core-like 267 GBM spheres incubated with or without proteinase K. n = 3 independent samples per group. b Venn diagram illustrating proteins identified by LC-MS/MS in CM from core-like and edge-like GBM spheres (left). Table showing the top 13 proteins exclusively detected in core-like CM (right). c Enzyme-linked immunosorbent assay (ELISA) for soluble CD109 in CM from 1051 and 101027 edge or core patient-derived GBM spheres. n = 2 independent experiments. d Graph demonstrating differences in the relative amount of proteins detected by LC-MS/MS in CM from core-like cells infected with shNT or with shHDAC1 encoding lentiviruses. e WB for HDAC1 and CD109 using 1051 core GBM cells infected with shNT or shHDAC1 lentiviruses or using 1051 edge GBM cells infected with GFP or HDAC1 encoding lentiviruses. f WB for CD109 and HDAC1 using core-like 267/1005 GBM spheres infected with shNT or shHDAC1 lentiviruses. g qRT-PCR analysis of CD109 expression in edge-like 157 GBM spheres infected with lentiviruses encoding GFP or HDAC1. n = 3 independent samples per group. h In vitro cell viability assay of edge-like 157 GBM spheres infected with lentiviruses encoding GFP or HDAC1 and subsequently irradiated (IR). n = 4 independent samples per group. i Flow cytometry analysis of caspase-3/7 activity and SYTOX staining in 1051 and 101027 edge or core spheres that were cultivated in a presence or absence of recombinant sCD109 for 3 days and subsequently irradiated with 8 Gy. j Bioluminescence imaging of mice intracranially co-injected with luciferase-labeled edge-like 157 GBM spheres and unlabeled shNT or shCD109-infected core-like 1005 GBM spheres (95:5 ratio) (left). Quantification of BLI signal in mice (right). n = 3 animals. k IF staining for GFP (green), c-Myc (red) and nucleus (blue) of mouse brains co-injected with GFP-labeled edge-like 157 GBM spheres and unlabeled shNT or shCD109-infected core-like 1005 GBM spheres. Scale bar 20 µm. a–j Data are mean ± s.d. Significance was calculated by unpaired, two-tailed t-test with **p < 0.01; ***p < 0.001.