human men1 sirna Search Results


sirna targeting men1  (OriGene)
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OriGene sirna targeting men1
Cell cycle subcellular localization of <t>menin</t> and MLL1-N. Immunofluorescence microscopy of HeLa cells stained for DNA, α-tubulin (α-Tub) and either (A) menin or (B) MLL1-N. Note that menin localizes to the mitotic spindle poles and mitotic spindle during early mitosis and to intercellular bridge microtubules during cytokinesis, similar to MLL1-N. Scale bars, 5 μm.
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Cell cycle subcellular localization of menin and MLL1-N. Immunofluorescence microscopy of HeLa cells stained for DNA, α-tubulin (α-Tub) and either (A) menin or (B) MLL1-N. Note that menin localizes to the mitotic spindle poles and mitotic spindle during early mitosis and to intercellular bridge microtubules during cytokinesis, similar to MLL1-N. Scale bars, 5 μm.

Journal: Endocrinology

Article Title: Menin Associates With the Mitotic Spindle and Is Important for Cell Division

doi: 10.1210/en.2019-00274

Figure Lengend Snippet: Cell cycle subcellular localization of menin and MLL1-N. Immunofluorescence microscopy of HeLa cells stained for DNA, α-tubulin (α-Tub) and either (A) menin or (B) MLL1-N. Note that menin localizes to the mitotic spindle poles and mitotic spindle during early mitosis and to intercellular bridge microtubules during cytokinesis, similar to MLL1-N. Scale bars, 5 μm.

Article Snippet: HeLa [CCL2; RRID:CVCL_0030 ( 22 ); ATCC] cell line growth and small interfering RNA (siRNA) treatments with OriGene control nontargeting siRNA (SR30004) and siRNA targeting MEN1 (SR302867A and SR302867B) were used as described previously ( 23 , 24 ).

Techniques: Immunofluorescence, Microscopy, Staining

Validation of menin’s localization to the mitotic spindle. (A) Immunoblot analysis showing that siMEN deplete menin protein levels compared with siCont. (B) Immunofluorescence microscopy of HeLa cells treated with siCont or siMEN for 48 h and stained for DNA, α-tubulin (α-Tub), and menin. Note that menin’s localization to the mitotic spindle is depleted in siMEN-treated cells. (C, D) Immunofluorescence microscopy of HeLa cells transfected with the overexpressed GFP-tagged version of menin (GFP-menin) and stained for (C) DNA, α-Tub, and MLL1-N, or (D) MLL1-C. Scale bars, 5 μm. Cont, control; MEN, menin; MW, molecular weight (kDa).

Journal: Endocrinology

Article Title: Menin Associates With the Mitotic Spindle and Is Important for Cell Division

doi: 10.1210/en.2019-00274

Figure Lengend Snippet: Validation of menin’s localization to the mitotic spindle. (A) Immunoblot analysis showing that siMEN deplete menin protein levels compared with siCont. (B) Immunofluorescence microscopy of HeLa cells treated with siCont or siMEN for 48 h and stained for DNA, α-tubulin (α-Tub), and menin. Note that menin’s localization to the mitotic spindle is depleted in siMEN-treated cells. (C, D) Immunofluorescence microscopy of HeLa cells transfected with the overexpressed GFP-tagged version of menin (GFP-menin) and stained for (C) DNA, α-Tub, and MLL1-N, or (D) MLL1-C. Scale bars, 5 μm. Cont, control; MEN, menin; MW, molecular weight (kDa).

Article Snippet: HeLa [CCL2; RRID:CVCL_0030 ( 22 ); ATCC] cell line growth and small interfering RNA (siRNA) treatments with OriGene control nontargeting siRNA (SR30004) and siRNA targeting MEN1 (SR302867A and SR302867B) were used as described previously ( 23 , 24 ).

Techniques: Biomarker Discovery, Western Blot, Immunofluorescence, Microscopy, Staining, Transfection, Control, Molecular Weight

Depletion of menin leads to cell division defects. (A) Immunofluorescence microscopy of HeLa cells treated with siCont or siMEN for 48 h and stained for DNA and α-tubulin (α-Tub). Note that siMEN cells show multiple aberrancies, including multipolar spindles and unaligned chromosomes in metaphase, lagging chromosomes in anaphase, multipolar cytokinesis, and multinucleated interphase cells. Arrows point to uncongressed chromosomes in a metaphase cell (panel with four arrows) and lagging chromosomes in a telophase cell (panel with one arrow). Scale bar, 5 μm. (B) Quantification of the percentage of mitotic cells with defective spindles, uncongressed chromosomes, and cytokinetic defects and interphase cells with more than one nucleus (multinucleated). Data represent the mean ± SD of three independent experiments, 100 cells counted for each. **P < 0.001. (C) Live-cell time-lapse microscopy snapshots of HCT116-GFP-H2B cells treated with siCont or siMEN (42). Representative cell division defects are shown, including cytokinetic arrest, multipolar cell division with cell death, and regression of a dividing cell into a binucleated cell. Time is in minutes. (D) The percentage of cells undergoing normal cell division, dying during cell division, undergoing aberrant cytokinesis and failing cytokinesis, and regressing to a binucleated state were quantified for siCont- or siMEN-treated cells. Data represent the mean ± SD of three independent experiments, 50 cells counted for each. **P < 0.001; ***P < 0.0001. (E) HeLa cells were treated with siCont or siMEN for 24 h, synchronized in G1/S with thymidine (Thy), and released into the cell cycle. Cells were harvested at the indicated time after release and extracts were immunoblotted for menin, cyclin B1 (CycB1), and α-Tub. Line graph shows relative cyclin B1 levels over time (in hours) normalized to α-Tub.

Journal: Endocrinology

Article Title: Menin Associates With the Mitotic Spindle and Is Important for Cell Division

doi: 10.1210/en.2019-00274

Figure Lengend Snippet: Depletion of menin leads to cell division defects. (A) Immunofluorescence microscopy of HeLa cells treated with siCont or siMEN for 48 h and stained for DNA and α-tubulin (α-Tub). Note that siMEN cells show multiple aberrancies, including multipolar spindles and unaligned chromosomes in metaphase, lagging chromosomes in anaphase, multipolar cytokinesis, and multinucleated interphase cells. Arrows point to uncongressed chromosomes in a metaphase cell (panel with four arrows) and lagging chromosomes in a telophase cell (panel with one arrow). Scale bar, 5 μm. (B) Quantification of the percentage of mitotic cells with defective spindles, uncongressed chromosomes, and cytokinetic defects and interphase cells with more than one nucleus (multinucleated). Data represent the mean ± SD of three independent experiments, 100 cells counted for each. **P < 0.001. (C) Live-cell time-lapse microscopy snapshots of HCT116-GFP-H2B cells treated with siCont or siMEN (42). Representative cell division defects are shown, including cytokinetic arrest, multipolar cell division with cell death, and regression of a dividing cell into a binucleated cell. Time is in minutes. (D) The percentage of cells undergoing normal cell division, dying during cell division, undergoing aberrant cytokinesis and failing cytokinesis, and regressing to a binucleated state were quantified for siCont- or siMEN-treated cells. Data represent the mean ± SD of three independent experiments, 50 cells counted for each. **P < 0.001; ***P < 0.0001. (E) HeLa cells were treated with siCont or siMEN for 24 h, synchronized in G1/S with thymidine (Thy), and released into the cell cycle. Cells were harvested at the indicated time after release and extracts were immunoblotted for menin, cyclin B1 (CycB1), and α-Tub. Line graph shows relative cyclin B1 levels over time (in hours) normalized to α-Tub.

Article Snippet: HeLa [CCL2; RRID:CVCL_0030 ( 22 ); ATCC] cell line growth and small interfering RNA (siRNA) treatments with OriGene control nontargeting siRNA (SR30004) and siRNA targeting MEN1 (SR302867A and SR302867B) were used as described previously ( 23 , 24 ).

Techniques: Immunofluorescence, Microscopy, Staining, Time-lapse Microscopy

Pharmacological inhibition of the menin-MLL1 interaction with MI-2 leads to cell division defects. (A) Immunofluorescence microscopy of HeLa cells treated with DMSO or MI-2 (10 μM) for 2 h before mitotic entry and stained for DNA and α-tubulin (α-Tub). Note that MI-2–treated cells show multiple aberrancies, including multipolar spindles and unaligned chromosomes in metaphase, lagging chromosomes in anaphase, multipolar cytokinesis, and multinucleated interphase cells. The arrow points to lagging chromosomes in a telophase cell. Scale bar, 5 μm. (B) Quantification of the percentage of mitotic cells with defective spindles, uncongressed chromosomes, and cytokinetic defects and interphase cells with more than one nucleus (multinucleated). Data represent the mean ± SD of three independent experiments, 100 cells counted for each. *P < 0.01; ***P < 0.0001. (C) Live-cell time-lapse microscopy snapshots of HCT116-GFP-H2B cells treated with DMSO or MI-2 (42). Representative cell division defects are shown, including multipolar cytokinesis and regression of dividing cells into binucleated cells. Time is in minutes. (D) The percentage of cells undergoing normal cell division, dying during cell division, undergoing defective divisions and failing cytokinesis, and regressing to a binucleated state were quantified for DMSO- or MI-2–treated cells. Data represent the mean ± SD of three independent experiments, 50 cells counted for each. **P < 0.001; ***P < 0.0001. (E) HeLa cells were synchronized in G1/S with thymidine (Thy) and released into the cell cycle in the presence of either DMSO or MI-2. Cells were harvested at the indicated time after release and extracts were immunoblotted for menin, cyclin B1 (CycB1), and α-Tub. Line graph shows relative cyclin B1 levels over time (in hours) normalized to α-Tub.

Journal: Endocrinology

Article Title: Menin Associates With the Mitotic Spindle and Is Important for Cell Division

doi: 10.1210/en.2019-00274

Figure Lengend Snippet: Pharmacological inhibition of the menin-MLL1 interaction with MI-2 leads to cell division defects. (A) Immunofluorescence microscopy of HeLa cells treated with DMSO or MI-2 (10 μM) for 2 h before mitotic entry and stained for DNA and α-tubulin (α-Tub). Note that MI-2–treated cells show multiple aberrancies, including multipolar spindles and unaligned chromosomes in metaphase, lagging chromosomes in anaphase, multipolar cytokinesis, and multinucleated interphase cells. The arrow points to lagging chromosomes in a telophase cell. Scale bar, 5 μm. (B) Quantification of the percentage of mitotic cells with defective spindles, uncongressed chromosomes, and cytokinetic defects and interphase cells with more than one nucleus (multinucleated). Data represent the mean ± SD of three independent experiments, 100 cells counted for each. *P < 0.01; ***P < 0.0001. (C) Live-cell time-lapse microscopy snapshots of HCT116-GFP-H2B cells treated with DMSO or MI-2 (42). Representative cell division defects are shown, including multipolar cytokinesis and regression of dividing cells into binucleated cells. Time is in minutes. (D) The percentage of cells undergoing normal cell division, dying during cell division, undergoing defective divisions and failing cytokinesis, and regressing to a binucleated state were quantified for DMSO- or MI-2–treated cells. Data represent the mean ± SD of three independent experiments, 50 cells counted for each. **P < 0.001; ***P < 0.0001. (E) HeLa cells were synchronized in G1/S with thymidine (Thy) and released into the cell cycle in the presence of either DMSO or MI-2. Cells were harvested at the indicated time after release and extracts were immunoblotted for menin, cyclin B1 (CycB1), and α-Tub. Line graph shows relative cyclin B1 levels over time (in hours) normalized to α-Tub.

Article Snippet: HeLa [CCL2; RRID:CVCL_0030 ( 22 ); ATCC] cell line growth and small interfering RNA (siRNA) treatments with OriGene control nontargeting siRNA (SR30004) and siRNA targeting MEN1 (SR302867A and SR302867B) were used as described previously ( 23 , 24 ).

Techniques: Inhibition, Immunofluorescence, Microscopy, Staining, Time-lapse Microscopy

Depletion of menin or inhibition of menin-MLL1 interaction does not perturb Kif2A localization. (A) Immunofluorescence microscopy of HeLa cells treated with siCont or siMEN for 48 h and stained for DNA and α-tubulin (α-Tub). Note that Kif2A remains localized to the mitotic spindle in siMEN-treated cells. Scale bar, 5 μm. (B, C) Immunofluorescence microscopy of HeLa cells treated with DMSO or MI-2 (10 μM) for 2 h before mitotic entry and stained for (B) DNA, α-Tub, and Kif2A, or (C) menin. Note that Kif2A remains localized to the mitotic spindle in MI-2–treated cells, whereas menin localization to the mitotic spindle decreases. Scale bar, 5 μm.

Journal: Endocrinology

Article Title: Menin Associates With the Mitotic Spindle and Is Important for Cell Division

doi: 10.1210/en.2019-00274

Figure Lengend Snippet: Depletion of menin or inhibition of menin-MLL1 interaction does not perturb Kif2A localization. (A) Immunofluorescence microscopy of HeLa cells treated with siCont or siMEN for 48 h and stained for DNA and α-tubulin (α-Tub). Note that Kif2A remains localized to the mitotic spindle in siMEN-treated cells. Scale bar, 5 μm. (B, C) Immunofluorescence microscopy of HeLa cells treated with DMSO or MI-2 (10 μM) for 2 h before mitotic entry and stained for (B) DNA, α-Tub, and Kif2A, or (C) menin. Note that Kif2A remains localized to the mitotic spindle in MI-2–treated cells, whereas menin localization to the mitotic spindle decreases. Scale bar, 5 μm.

Article Snippet: HeLa [CCL2; RRID:CVCL_0030 ( 22 ); ATCC] cell line growth and small interfering RNA (siRNA) treatments with OriGene control nontargeting siRNA (SR30004) and siRNA targeting MEN1 (SR302867A and SR302867B) were used as described previously ( 23 , 24 ).

Techniques: Inhibition, Immunofluorescence, Microscopy, Staining