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. 2022 May 26;10:869531. doi: 10.3389/fcell.2022.869531

FIGURE 1.

FIGURE 1

Characterization of nuclear F-actin structure in ovarian cancer cells. (A-B), OVCA432-nAc-citrine cells with nuclear F-actin structures. DAPI, blue. Green, nAc-citrine. A magnified image of the white rectangle is pictured in (B). Scale bar, 20 µm. (C), Proportion of cells with the presence of nuclear F-actin in ovarian cancer cell lines stably expressing nAc-citrine. The statistical analysis was performed using Graphpad Prism 9. The data was shown as mean ± s.e. ** p < 0.01 and *** p < 0.001.(D), Nuclear F-actin signal (green) within the nucleus of a representative cell of the OVCA432-nAc-citrine cell line obtained using confocal Z-stack scanning and ImageJ Reslice analysis. The bottom shows a cell without nuclear F-actin. Green, nAc-citrine. Blue, DAPI. Scale bar, 5 µm.(E), OVCA432-nAc-citrine cells stained for laminA/C (orange). Green, nAc-citrine. Blue, DAPI. Scale bar, 5 µm. (F), Nuclear F-actin in SKOV3 cells stained using rhodamine-phalloidin. SKOV3 cells were of no nAc-citrine expression. Scale bar, 10 µm. (G-H), Spheroids of SKOV3 cells stably expressing lifeact-citrine imaged after 4 days of spheroid growth. A magnified image of the white rectangle is pictured in (H). Green, lifeact-citrine. Blue, DAPI. Scale bar (in G), 20 µm. (I-J), F-actin staining for a frozen tumour section obtained from a high-grade serous ovarian cancer patient. Red, rhodamine-phalloidin. Blue, DAPI. The white rectangle area was magnified in J, showing a tumour cell stained for nuclear F-actin. Scale bar (in I), 10 µm. (K-L), Reslice algorithm of ImageJ was used to confirm the presence of nuclear F-actin within a patient tumour cell. K, Reslice analysis of a tumour cell (left panel) and rhodamine-phalloidin signal (red) in the nucleus (blue) (right panel). (L), negative control for Reslice analysis (left panel) and a cell without nuclear F-actin due to the absence of rhodamine-phalloidin signal (right panel). (M), Different views of an Imaris-modelled 3D image to visualize nuclear F-actin structures shown in J. Red, rhodamine-phalloidin. Blue, DAPI.