American Society of Hematology

Figure 2

$Figure 2. MMSET isoforms exhibit different patterns of nuclear distribution. (A) Distribution of MMSET isoforms in the chromatin and nuclear fractions. The full-length MMSET-II protein of the line KMS11 and all t(11;14) lines as well as the MB4-2 II variant of line H929 are chromatin-bound (lanes 1-2 and lanes 4-6, respectively), whereas the MB4-3 II variant of lines H929 and KMS12PE is additionally found in the soluble nuclear fraction (lanes 2 and 3). GAPDH is used as a loading control. CHR indicates insoluble chromatin fraction; NUC, soluble nuclear fraction. (B) Overexpressed MMSET protein is localized to nuclei and shows a diffuse pattern of expression. The t(4;14)-positive myeloma cell lines KMS11 and KMS28PE stained with anti-MMSET antibody show higher levels of MMSET expression (red) in the cell nucleus (blue) than the t(11;14) lines MM.M1 and KMS12BM. The full-length MMSET protein present in KMS11 exhibits a diffuse speckled nuclear distribution and is excluded from the nucleolus, whereas the MB4-3 protein present in KMS28PE appears focalized mainly in nucleoli. (C) NIH3T3 cells were transfected with an expression vector for MMSET-I or MMSET-II, immunostained with anti-MMSET antibody, and counterstained with phalloidin and DAPI. Original magnification ×400 for all panels.$

MMSET isoforms exhibit different patterns of nuclear distribution. (A) Distribution of MMSET isoforms in the chromatin and nuclear fractions. The full-length MMSET-II protein of the line KMS11 and all t(11;14) lines as well as the MB4-2 II variant of line H929 are chromatin-bound (lanes 1-2 and lanes 4-6, respectively), whereas the MB4-3 II variant of lines H929 and KMS12PE is additionally found in the soluble nuclear fraction (lanes 2 and 3). GAPDH is used as a loading control. CHR indicates insoluble chromatin fraction; NUC, soluble nuclear fraction. (B) Overexpressed MMSET protein is localized to nuclei and shows a diffuse pattern of expression. The t(4;14)-positive myeloma cell lines KMS11 and KMS28PE stained with anti-MMSET antibody show higher levels of MMSET expression (red) in the cell nucleus (blue) than the t(11;14) lines MM.M1 and KMS12BM. The full-length MMSET protein present in KMS11 exhibits a diffuse speckled nuclear distribution and is excluded from the nucleolus, whereas the MB4-3 protein present in KMS28PE appears focalized mainly in nucleoli. (C) NIH3T3 cells were transfected with an expression vector for MMSET-I or MMSET-II, immunostained with anti-MMSET antibody, and counterstained with phalloidin and DAPI. Original magnification ×400 for all panels.

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