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. 2021 Apr 30;11:663877. doi: 10.3389/fonc.2021.663877

Figure 5.

Figure 5

CCT2 overexpression promotes adherence of breast cancer cells in post-3D spheroid cultures and increases intracellular actin. (A) Brightfield and GFP overlay images of day 8 spheroids from T47D and MCF7 cells, CCT2-FLAG overexpressing and lentiviral control, transferred onto standard tissue culture plates are shown. After transfer, non-adherent cells were washed off and images were taken of remaining adherent cells. Adherent cells were then dissociated and counted by flow cytometry. Images were acquired using the Cytation 5 Cell Imaging Multi-Mode Reader (BioTek) (n=10). Magnification was 2.5X. (B) Confocal microscopy images of F-actin (stained with rhodamine phalloidin), DAPI, and overlays are shown for T47D, CCT2-FLAG overexpressing and lentiviral control, cells. Magnification was 40X. Inset shows a magnified view of cellular protrusions. To quantitate, integrated density was measured using ImageJ to calculate Corrected Total Cell Fluorescence (CTCF) as described in Methods. Values are mean with SD. *p-value <0.05, ****p-value <0.00005.