Figure 4
Setdb1 subcellular localisation changes upon terminal muscle differentiation and is dependent on Exportin-1. (a) Setdb1 localisation changes in C2C12 myoblasts during differentiation. Cells were proliferating (prolif.) or differentiating for 24 h (24 h diff.). Cellular Setdb1 (green) was detected by indirect IF. Cells were DAPI-stained to reveal DNA (red) prior to confocal fluorescent microscopy analyses. Scale bar=10 μm. (b) Quantification of Setdb1 localisation in proliferating (prolif.) and differentiating (24 h diff.) C2C12 myoblasts. Setdb1 localisation was classified as nuclear (N), homogeneous (H), or cytoplasmic (C). A minimum of 100 cells was counted for each condition. (c) Setdb1 delocalisation in differentiating C2C12 myoblasts is restricted by blocking nuclear export. Indirect IF of Setdb1 (green) and confocal microscopy were performed as described in (a). C2C12 myoblasts were differentiated for 24 h and non-treated (24 h diff.) or incubated in parallel with LMB for the last 18 h (24 h diff.+LMB). Scale bar=10 μm. (d) Quantification of Setdb1 localisation in C2C12 myoblasts differentiated for 24 h and non-treated (24 h diff.) or incubated in parallel with LMB for the last 18 h (24 h diff.+LMB). Setdb1 localisation was classified as described for (b). A minimum of 100 cells was counted for each condition. For (a, c): Images are representative of a minimum of three independent experiments. For (b, d): Data are presented as mean±s.e.m. of a minimum of three independent experiments. See also Supplementary Figure S4.
