Fig. 4.
The laforin/malin complex alters the subcellular localization of PKM2. a U2OS cells were transfected with plasmid pCMV-myc-PKM2. Cells were then treated or not with UV light (120 mJ/cm2; 120 s) to induce the accumulation of PKM2 into the nucleus. The localization of PKM2 was determined by immunofluorescence using anti-PKM2 antibodies (see Methods). DAPI staining was also performed to define the position of the nuclei. A merge image is presented. Scale bars are indicated. b U2OS cells expressing plasmid pCMV-myc-PKM2 were co-transfected with plasmids pFLAG-laforin and pCMV-HA-malin. Then, cells were treated of not with UV light as above. Panel shows the result in UV-treated cells. The localization of malin, a nuclear protein, and PKM2 was determined by immunofluorescence using anti-HA and anti-PKM2 antibodies, respectively. c The percentage of cells having a nuclear localization of PKM2 was determined in 100 cells from all the conditions. PKM-N: cells that present PKM2 inside the nucleus; PKM-C: cells that present PKM2 outside of the nucleus