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. 2021 Dec 25;9(12):e003973. doi: 10.1136/jitc-2021-003973

Figure 6.

Figure 6

ZEB1 regulate the formation of OvCa-TAMs spheroids. (A) The formation of SKOV3-TAMs spheroids, SKOV3oeZEB1-TAMs spheroids and SKOV3siZEB1-TAMs spheroids in the 3D coculture system. (B) The invasive and migratory capacity of SKOV3 isolated from SKOV3-TAMs spheroids, SKOV3oeZEB1-TAMs spheroids and SKOV3siZEB1-TAMs spheroids were measured by transwell-migration assay and scratch wound healing assay. (C) The quantitation of transwell-migration assay for isolated SKOV3 (n=4 biologically independent samples per group and an average of five fields acquired from each sample). (D) The quantitation of scratch wound healing assay for isolated SKOV3 (n=4 biologically independent samples per group and an average of five fields acquired from each sample). (E–H) qRT-PCR and Western bot analysis the gene and protein expression of E-CAD, SNAIL, TWIST, CCR8 and M-CSF. (I, J) The immunofluorescence staining for E-CAD, SNAIL, TWIST and CCR8. All Data were shown as mean±SD, and significant difference are indicated (*p<0.05, **p < 0.01 and ***p<0.001 against control. ###p < 0.001 against SKOV3 with the knock down of ZEB1 in spheroid). CCR8, Chemokine (C-C motif) ligand 18; 3D, three-dimensional; E-CAD, E-cadherin; M-CSF, macrophage colony-stimulating factor; OvCa, ovarian cancer; PBMC, peripheral blood monocyte cells; qRT-PCR, quantitative reverse transcription-PCR; TAMs, tumor-associated macrophages; ZEB1, zinc finger E-box-binding homeobox 1.