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. 2020 Mar 23;21(6):2203. doi: 10.3390/ijms21062203

Figure 2.

Figure 2

ERK5 requires SUMO modification for nuclear translocation. Immunofluorescence microscopy. (A) PC-3 cells were transfected with either GST-tagged ERK5 (wild type or K6/22R SUMO-deficient mutant) and MEK5DD or Cdc37 plasmids. After 24 h, cells were fixed with paraformaldehyde and immunofluorescent staining for ERK5 (green) using anti-GST antibody. Nuclei were stained with Hoechst (blue). (B) Subcellular fractionation. PC-3 cells were transfected with the indicated vectors and submitted to cellular fractionation as described in the Materials and Methods section. Proteins were detected by immunoblot using the indicated antibodies. (C) PC-3 cells were transfected with either GST-tagged ERK5 wild type (WT) or K6/22 mutant, left alone or treated with the nuclear export inhibitor leptomycin B (LMB, 20 nM) for 18 h and stained as in A. Similar results were obtained in three independent experiments. Scale bars, 10 μm.