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. 2016 Oct 14;113(44):E6813–E6822. doi: 10.1073/pnas.1608210113

Fig. S4.

Fig. S4.

Inhibition of either myosin II activity or actin polymerization increases FHL2 protein levels in the nucleus. (A) Western blot analysis for FHL2 with samples isolated from cytoplasmic or nuclear fraction of HFF cells treated with DMSO (control), blebbistatin (50 μM, 1 h), or cytochalasin D (2 μM, 0.5 h). (B, Top) SDS/PAGE with polyacrylamide containing Mn2+ and Phos-tag. Black arrow, phosphorylated FHL2; white arrow, unphosphorylated FHL2-GFP. (B, Bottom) SDS/PAGE with polyacrylamide, Western blot analysis for FHL2-GFP. Lane 1, cytoplasmic fraction in control cell; lane 2, nuclear fraction in control cell; lane 3, cytoplasmic fraction in the cell with blebbistatin treatment, lane 4, nuclear fraction in the cell with blebbistatin treatment; lane 5, cytoplasmic fraction in the cell with blebbistatin washout; lane 6, nuclear fraction in the cell with blebbistatin washout. HFF cells were treated with DMSO (control) or blebbistatin (50 μM, 1 h) or 1 h after washout of blebbistatin (washout). The cytoplasmic marker was α-tubulin; the nuclear marker was Lamin B1.