Journal: Cell reports

Article Title: A TAZ-AXL-ABL2 feed-forward signaling axis promotes lung adenocarcinoma brain metastasis

doi: 10.1016/j.celrep.2019.11.018

Figure Lengend Snippet: A) Representative images (day 30 post-injection) and B) analysis of brain metastasis-free survival (BMFS) in mice injected intracardially with PC9-pFuLT or pFuLT-Tet-TAZ4SA-expressing cells. Mice were given dox water for the duration of the study. Statistical analysis calculated by Log-rank (Mantel-Cox) test. Parental (n=13), TAZ4SA (n=17). C-D) Quantitative analysis (day 30 post-injection) of brain-metastatic index in mice injected intracardially with C) PC9-pFuLT (n=13) vs. PC9-pFuLT-Tet-TAZ4SA (n=17) or D) HCC4006-pFuLT (n=9) vs HCC4006-pFuLT-Tet-TAZ4SA (n=10) cells. Statistical analysis calculated by unpaired two-tailed t test. ** p-value < 0.01. E) Representative images of ex vivo mouse brains on day 32 post-injection with HCC4006-pFuLT or HCC4006-Tet-TAZ4SA cells. F) Diagram of iterative derivation of brain-metastatic cell lines. G) GSEA plot of TAZ4SA signature in PC9 parental vs PC9-BrM3 RNA-seq dataset. NES=normalized enrichment score. H) Venn diagram of overlapping transcripts upregulated in PC9-BrM3 and PC9-TAZ4SA cells. I) Representative images (day 42 post-injection) and J) BMFS in mice injected intracardially with PC9-BrM3 cells expressing non-target control (shNTC, n=10) or shTAZ (clone #73, n=10). K) Immunoblot to evaluate TAZ shRNA knockdown in PC9-BrM3 cells. Actin was used for protein loading control.

Article Snippet: Recombinant DNA & Plasmids pLKO-puro Non-Target shRNA Control Sigma Mission TRC1 SHC016-1EA pLKO-puro shAXL 1040 Sigma Mission TRC1 TRCN0000001040 pLKO-puro shTAZ 70 Sigma Mission TRC1 TRCN0000019470 pLKO-puro shTAZ 71 Sigma Mission TRC1 TRCN0000019471 pLKO-puro shTAZ 73 Sigma Mission TRC1 TRCN0000019473 TetO-FUW-pgk-puro ( Chowdhury et al., 2016 ) Addgene #85747; RRID:Addgene_85747 pLVX-Tight-Puro Tet-On Vector Xaralabos Varelas, Boston University, Boston, MA, USA N/A pLVX-TP-3F-TAZ4SA Xaralabos Varelas, Boston University, Boston, MA, USA N/A Scramble control shRNA ( Gu et al, 2016 ) N/A ABL1 shRNA ( Gu et al, 2016 ) N/A ABL2 shRNA ( Gu et al, 2016 ) N/A 3XFlag pCMV5-TOPO TAZ wild type ( Varelas et al., 2008 ) Addgene #24809; RRID:Addgene_24809 3XFlag pCMV5-TOPO TAZ Y321F This paper N/A Tet-pLKO-puro ( Wiederschain et al., 2009 ) Addgene #21915; RRID:Addgene_21915 Tet-pLKO-shNTC-puro This paper N/A Tet-pLKO-shAXL-puro This paper N/A, shAXL clone TRCN0000001040 Tet-pLKO-shTAZ-puro This paper N/A, shTAZ clone TRCN0000019470 N174-MCS http://n2t.net/addgene:81061 Addgene #81061; RRID:Addgene_81061 phL1A-pcDNA3 ( Hlavin and Lemmon, 1991 ) Addgene #12307; RRID:Addgene_12307 N174-L1CAM This paper N/A pN1-eGFP Anthony Koleske, Yale University, New Haven, CT, USA N/A pN1-ABL2-eGFP WT Anthony Koleske, Yale University, New Haven, CT, USA N/A pN1-ABL2-eGFP R198K This paper N/A pN1-ABL2-eGFP K317M This paper N/A pWZL Neo Myr Flag AXL ( Boehm et al., 2007 ) Addgene #20428; RRID:Addgene_20428 pWZL Neo Myr Flag AXL Y779F This paper N/A pWZL Neo Myr Flag AXL Y821F This paper N/A pWZL Neo Myr Flag AXL Y830F This paper N/A pWZL Neo Myr Flag AXL Y866F This paper N/A Software and Algorithms Prism 6 and 8 Graphpad http://graphpad.com/scientific-software/prism ImageJ ( Schneider et al., 2012 ) http://imagej.nih.gov RStudio R Foundation for Statistical Computing http://rstudio.com CBLigand Online BBB Predictor Tool v0.90 ( Liu et al., 2014 ) http://cbligand.org Living Image Perkin Elmer http://perkinelmer.com GSEA 3.0 ( Subramanian et al., 2005 ) http://software.broadinstitute.org/gsea KM Plotter ( Gyorffy et al., 2013 ) http://kmplot.com CBioPortal v3.1.1 ( Cerami et al., 2012 ; Gao et al., 2013 ) http://cbioportal.org TrimGalore v0.4.4 Babraham Bioinformatics http://github.com/FelixKrueger/TrimGalore STAR aligner ( Dobin et al., 2013 ) http://github.com/alexdobin/STAR HTSeq-count ( Anders et al., 2015 ) http://htseq.readthedocs.io DESeq2 v2.12 ( Love et al., 2014 ) http://bioconductor.org/packages/release/bioc/html/DESeq2.html Hisat2 v2.1.0 ( Kim et al., 2019 ) http://ccb.jhu.edu/software/hisat2/index.shtml Open in a separate window KEY RESOURCES TABLE Co-immunoprecipitation assays.

Techniques: Injection, Expressing, Two Tailed Test, Ex Vivo, RNA Sequencing, Control, Western Blot, shRNA, Knockdown

Journal: Cell reports

Article Title: A TAZ-AXL-ABL2 feed-forward signaling axis promotes lung adenocarcinoma brain metastasis

doi: 10.1016/j.celrep.2019.11.018

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Recombinant DNA & Plasmids pLKO-puro Non-Target shRNA Control Sigma Mission TRC1 SHC016-1EA pLKO-puro shAXL 1040 Sigma Mission TRC1 TRCN0000001040 pLKO-puro shTAZ 70 Sigma Mission TRC1 TRCN0000019470 pLKO-puro shTAZ 71 Sigma Mission TRC1 TRCN0000019471 pLKO-puro shTAZ 73 Sigma Mission TRC1 TRCN0000019473 TetO-FUW-pgk-puro ( Chowdhury et al., 2016 ) Addgene #85747; RRID:Addgene_85747 pLVX-Tight-Puro Tet-On Vector Xaralabos Varelas, Boston University, Boston, MA, USA N/A pLVX-TP-3F-TAZ4SA Xaralabos Varelas, Boston University, Boston, MA, USA N/A Scramble control shRNA ( Gu et al, 2016 ) N/A ABL1 shRNA ( Gu et al, 2016 ) N/A ABL2 shRNA ( Gu et al, 2016 ) N/A 3XFlag pCMV5-TOPO TAZ wild type ( Varelas et al., 2008 ) Addgene #24809; RRID:Addgene_24809 3XFlag pCMV5-TOPO TAZ Y321F This paper N/A Tet-pLKO-puro ( Wiederschain et al., 2009 ) Addgene #21915; RRID:Addgene_21915 Tet-pLKO-shNTC-puro This paper N/A Tet-pLKO-shAXL-puro This paper N/A, shAXL clone TRCN0000001040 Tet-pLKO-shTAZ-puro This paper N/A, shTAZ clone TRCN0000019470 N174-MCS http://n2t.net/addgene:81061 Addgene #81061; RRID:Addgene_81061 phL1A-pcDNA3 ( Hlavin and Lemmon, 1991 ) Addgene #12307; RRID:Addgene_12307 N174-L1CAM This paper N/A pN1-eGFP Anthony Koleske, Yale University, New Haven, CT, USA N/A pN1-ABL2-eGFP WT Anthony Koleske, Yale University, New Haven, CT, USA N/A pN1-ABL2-eGFP R198K This paper N/A pN1-ABL2-eGFP K317M This paper N/A pWZL Neo Myr Flag AXL ( Boehm et al., 2007 ) Addgene #20428; RRID:Addgene_20428 pWZL Neo Myr Flag AXL Y779F This paper N/A pWZL Neo Myr Flag AXL Y821F This paper N/A pWZL Neo Myr Flag AXL Y830F This paper N/A pWZL Neo Myr Flag AXL Y866F This paper N/A Software and Algorithms Prism 6 and 8 Graphpad http://graphpad.com/scientific-software/prism ImageJ ( Schneider et al., 2012 ) http://imagej.nih.gov RStudio R Foundation for Statistical Computing http://rstudio.com CBLigand Online BBB Predictor Tool v0.90 ( Liu et al., 2014 ) http://cbligand.org Living Image Perkin Elmer http://perkinelmer.com GSEA 3.0 ( Subramanian et al., 2005 ) http://software.broadinstitute.org/gsea KM Plotter ( Gyorffy et al., 2013 ) http://kmplot.com CBioPortal v3.1.1 ( Cerami et al., 2012 ; Gao et al., 2013 ) http://cbioportal.org TrimGalore v0.4.4 Babraham Bioinformatics http://github.com/FelixKrueger/TrimGalore STAR aligner ( Dobin et al., 2013 ) http://github.com/alexdobin/STAR HTSeq-count ( Anders et al., 2015 ) http://htseq.readthedocs.io DESeq2 v2.12 ( Love et al., 2014 ) http://bioconductor.org/packages/release/bioc/html/DESeq2.html Hisat2 v2.1.0 ( Kim et al., 2019 ) http://ccb.jhu.edu/software/hisat2/index.shtml Open in a separate window KEY RESOURCES TABLE Co-immunoprecipitation assays.

Techniques: Western Blot, Immunofluorescence, Recombinant, In Vivo, In Vitro, Mutagenesis, Caspase-Glo Assay, RNA Sequencing, shRNA, Control, Plasmid Preparation, Software

Journal: eLife

Article Title: TAZ inhibits glucocorticoid receptor and coordinates hepatic glucose homeostasis in normal physiological states

doi: 10.7554/eLife.57462

Figure Lengend Snippet:

Article Snippet: Recombinant DNA reagent , pCMV-TOPO TAZΔWW (human) , Addgene , Cat. #: 24811RRID: Addgene_24811 , PMID: 18568018.

Techniques: Blocking Assay, Plasmid Preparation, Expressing, Enzyme-linked Immunosorbent Assay, Reporter Assay, Mutagenesis, cDNA Synthesis, Extraction, Glucose Oxidase Assay, Recombinant, Construct, Control, Knockdown, Generated, Sequencing, Cell Culture, In Vivo, Protease Inhibitor, SYBR Green Assay, Membrane, Transfection

Journal: Cell

Article Title: Inflammation Improves Glucose Homeostasis Through IKKβ-XBP1s Interaction

doi: 10.1016/j.cell.2016.10.015

Figure Lengend Snippet: (A) XBP1s and IKKβ levels in HEK293 cells infected with XBP1s-expressing adenovirus (Ad-XBP1s) and different doses of constitutively active IKKβ expressing adenovirus (Ad-ca-IKKβ).

Article Snippet: Plasmid: Human XBP1s plasmid , Addgene , Cat#63680.

Techniques: Infection, Expressing

Journal: Cell

Article Title: Inflammation Improves Glucose Homeostasis Through IKKβ-XBP1s Interaction

doi: 10.1016/j.cell.2016.10.015

Figure Lengend Snippet: (A) Immunoblotting for HA-tagged ca-IKKβ and flag-tagged XBP1s proteins after immunoprecipitation of XBP1s from HEK293 cells.

Article Snippet: Plasmid: Human XBP1s plasmid , Addgene , Cat#63680.

Techniques: Western Blot, Immunoprecipitation

Journal: Cell

Article Title: Inflammation Improves Glucose Homeostasis Through IKKβ-XBP1s Interaction

doi: 10.1016/j.cell.2016.10.015

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Plasmid: Human XBP1s plasmid , Addgene , Cat#63680.

Techniques: Recombinant, Plasmid Preparation, Expressing, Sequencing, Software