Cooperative high expression of NSD2, SMARCA5, and BAZ1B promotes PCH localization of H3K36me2. (A) Relative protein levels of NSD2, SMARCA5, and BAZ1B in mouse cell lines evaluated by western blotting. Lamin B was detected as a loading control. The positions of size standards are indicated on the left. (B) PCH-to-nucleus intensity ratio of H3K36me2. Pmi28 cells were transfected with the expression vectors of indicated proteins, fixed, and stained with anti-H3K36me2, JF646 HaloTag ligand, and Hoechst 33342. When expressing three proteins, an equal amount of each expression vector was mixed. When expressing one or two proteins, the same amount of each vector was used as was the case of triple expression, and the total DNA amount was adjusted using the EGFP- or HaloTag-expression vector. See Fig. 1 legend for the details of box plots. Note that P values were calculated using the Mann–Whitney U test and Hommel correction with the EGFP control. Source data are available for this figure: SourceData F7.