ZFIN Image: Darp et al., 2022, Fig. 6

Image

Figure Caption

Fig. 6

Supernumerary centrioles contribute to RhoA downregulation and BRAF^V600E-induced tetraploidy.

A DAPI, anti-CENTRIN-2 and anti-RHOA staining in control (?Dox) and BRAF^V600E-expressing (+Dox) anaphase RPE-1 cells. Insets show centrioles at one pole. Images are maximum intensity projections of z-stacks. Scale bar = 7.5?�M. B Mean RHOA fluorescence intensity at the equator of control RPE-1 anaphase cells (n?=?30) and BRAF^V600E-expressing RPE-1 anaphase cells with normal (4) and supernumerary (>4) centrosomes. N?=?41 cells for ?Dox, N?=?35 for +Dox with normal centrioles, and N?=?24 for +Dox with supernumerary centrioles. Brown-Forsythe and Welch one-way ANOVA with Dunnett?s multiple comparisons test. Error bars represent mean?�?SEM. C Percent RPE-1 cells in mitosis with supernumerary (>4) centrioles in control RPE-1 cells, BRAFV600E-expressing RPE-1 cells and BRAFV600E-expressing RPE-1 cells treated with Centrinone. Centrinone was added coincident with Dox administration. N?=?3 independent experiments examining ?Dox = 119, +Dox = 124, +Dox +Centrinone = 122 cells. One-way ANOVA with Tukey?s multiple comparisons test. Error bars represent mean?�?SEM. D DAPI and anti-RHOA staining in -BRAF^V600E (?Dox) RPE-1 cells, BRAF^V600E-expressing (+Dox) RPE-1 cells, and BRAF^V600E-expressing (+Dox) RPE-1 cells treated with Centrinone. Centrinone was added coincident with Dox administration (G1/S/G2/M) or only during G1/S. Images are maximum intensity projections of z-stacks (0.20?�M). Scale bar = 7.5?�M. E Mean RHOA fluorescence intensity at the equator of control RPE-1 cells, BRAF^V600E-expressing RPE-1 cells, and BRAF^V600E-expressing RPE-1 cells treated with Centrinone coincident with Dox administration (G1/S/G2/M) or only during G1/S. N?=?41 cells for ?Dox, N?=?39 for +Dox, N?=?44 for +Dox +Centrinone in G1/S/G2/M, and N?=?35 for +Dox +Centrinone in G1/S. Brown-Forsythe and Welch one-way ANOVA with Dunnett?s multiple comparisons test. Error bars represent mean?�?SEM. F Fold change in G1 tetraploids in control RPE-1 FUCCI cells (?Dox), BRAF^V600E-expressing RPE-1 FUCCI cells (+Dox) and BRAF^V600E-expressing RPE-1 FUCCI cells treated with Centrinone (+Dox +cent.). Fold changes are expressed relative to the control cells. N?=?3 independent experiments. One-way ANOVA with Tukey?s multiple comparisons test. Error bars represent mean?�?SEM.

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Nat. Commun.