In vivo and in vitro immunostaining of trimethylated histone H3 in P. misakiensis. (A-D)Growing buds stained with anti-H3K27me3 antibody. (B, D)DAPI staining after immunostaining. (A, B)Bars, 50 μm. (C, D)Bars, 25 μm. Black and white arrowheads show nuclei of morula cells. (E-G)Cultured cells stained with anti-H3K27me3 antibody. (E)Cells not treated with TC14-3. Bar, 50 μm. (F)Cells treated with TC14-3E106G. (G)Cells treated with wild-type TC14-3. Bar, 50 μm. (H)Western blotting of cell lysates. Lanes 1, anti-histone H3 antibody staining. Arrowhead shows histone H3. Lanes 2-4, anti-H3K27me3 antibody staining. Lane 2, cells not treated with TC14-3. Lane 3, cells treated with TC14-3E106G. Lane 4, cells treated with wild-type TC14-3. ae, atrial epithelium; e, epidermis; h, hemocoel; m, morula cell.