Journal: Cell reports

Article Title: Sensing Actin Dynamics through Adherens Junctions

doi: 10.1016/j.celrep.2020.01.106

Figure Lengend Snippet: (A–D) Projections of all x-y optical slices of A431 cells stained for (A) E-cadherin (Ecad, red) and F-actin (actin, green); (B) Ecad (green), F-actin (red), and myosin IIA (myosin II, blue); (C) Ecad (green), actin, and calponin-3 (Cnn3); and (D) Cnn3 (green) and myosin IIA (red).

Article Snippet: The plasmid encoding PA-mCH-tagged b-actin was a gift from Dr. Verkhusha (Addgene, plasmid # 31949). pRc-CNN3-mGFP plasmid was generated by using Myc-DDK tagged CNN3 (Origene, RC200488).

Techniques: Staining

Journal: Cell reports

Article Title: Sensing Actin Dynamics through Adherens Junctions

doi: 10.1016/j.celrep.2020.01.106

Figure Lengend Snippet: (A–D) Projections of all x-y optical slices of Cnn3-Dn-expressing A431 cells imaged for Dendra2 fluorescence (green, Cnn3-Dn) and stained for (A) Ecad, (B) actin, (C) vinculin (Vcn), and (D) mena. Scale bar, 10 μm. The zoomed areas (indicated by dashed boxes in Cnn3-Dn images) are presented at the bottom. Scale bar, 5 μm. Arrowheads indicate the gaps in Cnn3-Dn distribution along pAJ-linked bundles that are precisely matched by the antibody staining.

Article Snippet: The plasmid encoding PA-mCH-tagged b-actin was a gift from Dr. Verkhusha (Addgene, plasmid # 31949). pRc-CNN3-mGFP plasmid was generated by using Myc-DDK tagged CNN3 (Origene, RC200488).

Techniques: Expressing, Fluorescence, Staining

Journal: Cell reports

Article Title: Sensing Actin Dynamics through Adherens Junctions

doi: 10.1016/j.celrep.2020.01.106

Figure Lengend Snippet: (A–C) Maximum projection view of control A431-EcGFP cells (A431-EcGFP) or after Cfl1-KO (A431-EcGFP-Cfl1-KO). Cells were imaged for GFP (EcGFP) and for (A) Cfl1 (Cfl, red), (B) actin (actin, red), and (C) Cnn-3 (Cnn3, red). Scale bar, 10 μm. The zoomed areas (indicated by dashed boxes in EcGFP images) are presented in the insets (A) or at the bottom (B and C). Scale bar, 5 μm.

Article Snippet: The plasmid encoding PA-mCH-tagged b-actin was a gift from Dr. Verkhusha (Addgene, plasmid # 31949). pRc-CNN3-mGFP plasmid was generated by using Myc-DDK tagged CNN3 (Origene, RC200488).

Techniques:

Journal: Cell reports

Article Title: Sensing Actin Dynamics through Adherens Junctions

doi: 10.1016/j.celrep.2020.01.106

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: The plasmid encoding PA-mCH-tagged b-actin was a gift from Dr. Verkhusha (Addgene, plasmid # 31949). pRc-CNN3-mGFP plasmid was generated by using Myc-DDK tagged CNN3 (Origene, RC200488).

Techniques: Recombinant, CRISPR, Plasmid Preparation, Software

Journal: Cell Death & Disease

Article Title: MEIS2 is essential for neuroblastoma cell survival and proliferation by transcriptional control of M-phase progression

doi: 10.1038/cddis.2014.370

Figure Lengend Snippet: MEIS2 is essential for neuroblastoma cell survival. ( a ) Immunoblotting of MEIS2 in a panel of 13 human neuroblastoma cell lines. β -Actin levels are shown as a loading control. ( b and c ) qRT-PCR ( b ) and immunoblot ( c ) analyses of MEIS2 expression in BE(2)-C cells infected with lentiviruses expressing either shGFP or shMEIS2. Error bars ( b ) represent S.D. ( n =3). GAPDH ( c ) serves as a loading control. ( d ) Micrographs of BE(2)-C cells infected for 3 days with lentiviruses expressing either shGFP or shMEIS2. ( e ) Trypan blue exclusion assay of viable BE(2)-C cells infected for 4 days with lentiviruses expressing either shGFP or shMEIS2

Article Snippet: Immunoblotting was conducted according to standard procedures using the following primary antibodies: rabbit anti-BCL2 (sc-783, 1 : 200), rabbit anti-caspase 3 (sc-7148, 1 : 200), rabbit anti-Flag (F7425, 1 : 1000; Sigma-Aldrich), rabbit anti-FOXM1 (sc-502, 1 : 100), rabbit anti-GAPDH (sc-25778, 1 : 3000), mouse anti-MEIS2 (sc-81986, 1 : 400), rabbit anti- β -actin (600-401-886, 1 : 2000; Rockland Immunochemicals, Gilbertsville, PA, USA), and mouse anti- α -tubulin (B-5-1-2, 1 : 5000; Sigma-Aldrich).

Techniques: Western Blot, Quantitative RT-PCR, Expressing, Infection, Trypan Blue Exclusion Assay

Journal: Cell Death & Disease

Article Title: MEIS2 is essential for neuroblastoma cell survival and proliferation by transcriptional control of M-phase progression

doi: 10.1038/cddis.2014.370

Figure Lengend Snippet: MEIS2 targets the MuvB-BMYB-FOXM1 complex for transcriptional control of M-phase progression. ( a ) qRT-PCR analysis of mRNA expression of MEIS2, FOXM1, BMYB, and RBBP4 in BE(2)-C cells infected with lentiviruses expressing either shGFP or shMEIS2. ( b ) Immunoblotting of MEIS2 and FOXM1 in BE(2)-C cells infected with lentiviruses expressing either shGFP or shMEIS2. β -Actin levels are shown as a loading control. MEIS2 and FOXM1 levels were quantified against β -actin. ( c ) GSEA showing marked downregulation of the FOXM1 pathway genes in BE(2)-C cells with MEIS2 depletion. ( d ) ChIP-qPCR analysis showing MEIS2 binding to the FOXM1 promoter region (−312 to −158) containing a consensus TGIF/MEIS2-binding sequence TGTCA. Error bars, S.D. ( n =3). ( e ) Immunoblotting of FOXM1 in BE(2)-C and SK-N-DZ cells with or without FOXM1 knockdown. β -Actin levels are shown as a loading control. ( f ) qRT-PCR analysis of mRNA expression of key FOXM1 target genes in BE(2)-C and SK-N-DZ cells with or without FOXM1 knockdown. ( g ) Growth assay of BE(2)-C and SK-N-DZ cells with or without FOXM1 knockdown. Error bars, S.D. ( n =3). ** P <0.001. Data ( d and g ) were analyzed with two-tailed Student's t -test

Article Snippet: Immunoblotting was conducted according to standard procedures using the following primary antibodies: rabbit anti-BCL2 (sc-783, 1 : 200), rabbit anti-caspase 3 (sc-7148, 1 : 200), rabbit anti-Flag (F7425, 1 : 1000; Sigma-Aldrich), rabbit anti-FOXM1 (sc-502, 1 : 100), rabbit anti-GAPDH (sc-25778, 1 : 3000), mouse anti-MEIS2 (sc-81986, 1 : 400), rabbit anti- β -actin (600-401-886, 1 : 2000; Rockland Immunochemicals, Gilbertsville, PA, USA), and mouse anti- α -tubulin (B-5-1-2, 1 : 5000; Sigma-Aldrich).

Techniques: Quantitative RT-PCR, Expressing, Infection, Western Blot, Binding Assay, Sequencing, Growth Assay, Two Tailed Test