American Society of Hematology

Figure 1

$Figure 1. Validation of the T26 mAb. (A) Western blotting analysis of cells expressing or not the NPM1mutA protein. Lysates from the indicated cell lines were analyzed using mAbs specific to NPM1mutA (T26), wt NPM1 (NPMc), or tubulin β (Santa Cruz Clone H-235). (Left panel) Total lysates (T) were fractionated into cytoplasmic (C) or nuclear (N) fractions. (Right panel) Lysates were prepared from not infected OCI-AML3 cells (NI) or the same cells after infection with lentiviral vector expressing (shNPM1mutA) or not (empty vector; EV) shRNAs against NPM1mutA. (B) IF analysis of the indicated cell lines with the specified antibodies (T26 or NPMc). Staining with T26 or NPMc, DAPI staining, and merged channels (Merge) are shown. Original magnifications: left panels, ×600; right panels, ×1000. (C) IHC of bone marrow trephine biopsy sections from NPMc+ AML (left) or wt NPM AML (right) patients.$

Validation of the T26 mAb. (A) Western blotting analysis of cells expressing or not the NPM1mutA protein. Lysates from the indicated cell lines were analyzed using mAbs specific to NPM1mutA (T26), wt NPM1 (NPMc), or tubulin β (Santa Cruz Clone H-235). (Left panel) Total lysates (T) were fractionated into cytoplasmic (C) or nuclear (N) fractions. (Right panel) Lysates were prepared from not infected OCI-AML3 cells (NI) or the same cells after infection with lentiviral vector expressing (shNPM1mutA) or not (empty vector; EV) shRNAs against NPM1mutA. (B) IF analysis of the indicated cell lines with the specified antibodies (T26 or NPMc). Staining with T26 or NPMc, DAPI staining, and merged channels (Merge) are shown. Original magnifications: left panels, ×600; right panels, ×1000. (C) IHC of bone marrow trephine biopsy sections from NPMc⁺ AML (left) or wt NPM AML (right) patients.

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