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. 2021 Sep 4;24(9):103086. doi: 10.1016/j.isci.2021.103086

Figure 2.

Figure 2

The DDX3-AREG axis promotes cell migration, angiogenesis, and macrophage differentiation in vitro

(A) SAS cells were transfected with pP2A (vec) or pP2A-AREG for 72 hours. Immunoblotting of the cell lysates, cell morphology, and percentage of spindle cells are shown (mean ± SD; ∗p<0.01). Scale bars, 50 μm.

(B) Boyden chamber assay of SAS cells in DMEM or CM derived from control (shC) or DDX3-depleted (shD) SAS cells supplemented with or without 5 ng/ml rAREG (mean ± SD; ∗p<0.01). Immunoblotting was performed in shRNA-transfected SAS cells. Scale bar, 500 μm.

(C) Angiogenesis assay of EA.hy926 cells that were cultured in DMEM or CM as in (B). Bar graph quantifies tube formation by EA.hy926 cells (mean ± SD; ∗p<0.01). Scale bars, 500 μm.

(D) RT-qPCR analysis of the indicated transcripts in THP-1 cells cultured in DMEM or the various CM, as indicated (mean ± SD; ∗p<0.01).

(E) Schematic diagram shows experimental procedures for collection of CM used for angiogenesis assays. EA.hy926 cells were cultured in CM from THP-1 that had been cultured in CM from siC or siD-transfected SAS cells. Bar graph quantifies tube formation by EA.hy926 cells (mean ± SD; ∗p<0.01). Scale bar, 500 μm.