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Fig. 2 | Genome Biology

Fig. 2

From: Transcriptional regulatory networks of tumor-associated macrophages that drive malignancy in mesenchymal glioblastoma

Fig. 2

MARCO promotes mesenchymal, invasive, and migratory phenotypes, as well as therapeutic resistance to irradiation. a qRT-PCR analysis to determine the effects of Pparg on the mRNA expression levels of the MA-TAM genes Marco, Ccl7, Fpr3, and Areg in mouse peritoneal macrophages. b Representative immunofluorescence images of Pparg and Marco in mouse peritoneal macrophages transduced with either control or Pparg. c qRT-PCR analysis to determine the effects of siPPARG, siBatf, or siSpi1 on the mRNA expression levels of the MA-TAM genes Marco, Ccl7, Cd180, Cr2, Mmp8, and Areg in mouse peritoneal macrophages. d qRT-PCR analysis to determine the effects of MARCO or CCL7 on the expression of mesenchymal, stemness, and cellular invasion markers including CD44, NANOG, LIF, and MMP-2 in two GSC samples. e Immunoblot analysis of CD44, TAZ, and NANOG activity in GSCs treated with either control, MARCO, or CCL7; α-tubulin was used as a loading control. f Representative images of 3D invasion assays in GSCs treated with either control, monocyte-derived CM, MARCOlow TAM-derived CM, or MARCOhigh TAM-derived CM (upper panel). Immunofluorescence images of CD44 intracellular domain and DAPI (red and blue, respectively; lower panel). g Representative images of tube formation assay (left panel) and representative bar graphs (right panel) for HUVECs. h The 1/stem cell frequency of GSCs treated with either monocyte-derived CM, MARCOlow TAM-derived CM, or MARCOhigh TAM-derived CM and subjected to 2 Gy ionizing radiation. Stem cell frequency was calculated by extreme limiting dilution analysis. i qRT-PCR analysis to determine the effects of monocyte-derived CM, MARCOlow TAM-derived CM, and MARCOhigh TAM-derived CM on mesenchymal, stemness, and cellular invasion markers in GSCs. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001. Data shown in a, c, d, and i are representative of three independent and reproducible experiments. Data shown in b and eh are representative of two independent and reproducible experiments

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