Characterization of the farnesylation-deficient LKB1C433S/C433S mouse (A) The knockin construct, the endogenous LKB1 allele containing exons 1–10 and the.

Slides:

Advertisements

Similar presentations

Supplementary Figure 1. Generation of hepatocyte-specific A20 knockout mice. Targeting scheme. Diagram showing the LoxP-flanked (Floxed) and the deleted.

Advertisements

Guaiacol Treatment Decreases Resting Glycogen Synthase Activity and Expands the Life Span in the Mouse Models of APBD H. Orhan AKMAN1, Yasemin G. Kurt1,

A conditional knockout mouse model reveals endothelial cells as the principal and possibly exclusive source of plasma factor VIII by Scot A. Fahs, Matthew.

Tamoxifen induced deletion of FAK

Vascular Endothelial-Specific Dimethylarginine Dimethylaminohydrolase-1–Deficient Mice Reveal That Vascular Endothelium Plays an Important Role in Removing.

Volume 109, Issue 1, Pages (April 2002)

Volume 24, Issue 1, Pages (January 2006)

Supplementary Figure S12

Differentiation of CD4+ T Cells to Th1 Cells Requires MAP Kinase JNK2

ADAMTS-5 deficient mice do not develop mechanical allodynia associated with osteoarthritis following medial meniscal destabilization A.M. Malfait, J.

Important scaffold function of the Janus kinase 2 uncovered by a novel mouse model harboring a Jak2 activation-loop mutation by Eric Keil, David Finkenstädt,

Christoph Heiner Westphal, Philip Leder Current Biology

by Hiromi Gunshin, Carolyn N. Starr, Cristina DiRenzo, Mark D

by Yuko Kimura, Takashi Miwa, Lin Zhou, and Wen-Chao Song

Volume 2, Issue 4, Pages (October 1998)

Lack of Enhanced Spinal Regeneration in Nogo-Deficient Mice

Volume 19, Issue 6, Pages (December 2003)

Volume 9, Issue 3, Pages (September 1998)

Volume 154, Issue 6, Pages (September 2013)

RBM24 Is a Major Regulator of Muscle-Specific Alternative Splicing

Polina Iakova, Samir S Awad, Nikolai A Timchenko Cell

Generation of γ1 EQ conditional knock-in mice.

Volume 14, Issue 1, Pages (January 2008)

Volume 13, Issue 22, Pages (November 2003)

Volume 11, Issue 4, Pages (October 2006)

Volume 18, Issue 2, Pages (February 2003)

Volume 12, Issue 5, Pages (November 2010)

Volume 15, Issue 6, Pages (March 2005)

Molecular Therapy - Nucleic Acids

Naokazu Inoue, Ph. D. , Takao Nishikawa, M. S. , Masahito Ikawa, Ph. D

Genetic disruption of p38α Tyr323 phosphorylation prevents T-cell receptor–mediated p38α activation and impairs interferon-γ production by Ludmila Jirmanova,

Volume 88, Issue 4, Pages (February 1997)

Volume 6, Issue 3, Pages (March 1997)

Volume 127, Issue 1, Pages (October 2006)

WRKY20 dual sgRNA approach.

Volume 13, Issue 9, Pages (December 2015)

Volume 19, Issue 5, Pages (November 2003)

Role of the INK4a Locus in Tumor Suppression and Cell Mortality

Toyoaki Natsume, Tomomi Kiyomitsu, Yumiko Saga, Masato T. Kanemaki

Volume 92, Issue 4, Pages (February 1998)

Molecular Therapy - Nucleic Acids

Targeted Mutagenesis of the Endogenous Mouse Mis Gene Promoter

The LKB1 tumor suppressor negatively regulates mTOR signaling

APOE Gene Targeting (A) Schematic representation of the endogenous APOE locus, the gene targeting vector and the targeted APOE locus. The exons of the.

Volume 10, Issue 4, Pages (April 1999)

Generation of Siva conditional knockout mice.

Volume 17, Issue 2, Pages (August 2015)

Hua Gao, Yue Sun, Yalan Wu, Bing Luan, Yaya Wang, Bin Qu, Gang Pei

Deletion of Srf with a skeletal muscle-specific Cre transgene.

Mice with AS160/TBC1D4-Thr649Ala Knockin Mutation Are Glucose Intolerant with Reduced Insulin Sensitivity and Altered GLUT4 Trafficking Shuai Chen, David.

Fig. 4. Targeted disruption of STK35 transcripts in mouse.

Volume 6, Issue 4, Pages (October 2007)

Fig. 1. Generation of WNK3 knockout mice

Detection of gene recombination in endoxifen- or Adeno-Cre-induced tumours. Detection of gene recombination in endoxifen- or Adeno-Cre-induced tumours.

Volume 17, Issue 4, Pages (October 2002)

14-3-3γ phosphorylated at S59 by Lats2 in response to UV damage.

Modulation of insulin sensitivity by IL-6 in mice: A lack of PTP1B prevents chronic effects of IL-6. Modulation of insulin sensitivity by IL-6 in mice:

Gα12 is not required for hepatic cystogenosis induced by Pkd1 knockout

Full-length TAPL interacts specifically with LAMP-1 and LAMP-2.

Generation and genotyping of Syk-AQL mice.

Posttranslational Mechanisms Regulate the Mammalian Circadian Clock

Targeted disruption of the mouse Atp1a2.

Volume 14, Issue 1, Pages (January 2001)

DAPK interacts with HSF1 in vivo and in vitro.

Volume 7, Issue 3, Pages (March 2005)

A CDK-Independent Function of Mammalian Cks1

Key functional sites of SPINDLIN1 could be phosphorylated by Aurora-A.

CTCF regulates cell cycle progression of αβ T cells in the thymus

Analysis of Type 2 Immunity In Vivo with a Bicistronic IL-4 Reporter

Keratinocyte-specific disruption of the Stat3 gene by the Cre-LoxP system. Keratinocyte-specific disruption of the Stat3 gene by the Cre-LoxP system. A,

Presentation transcript:

Characterization of the farnesylation-deficient LKB1C433S/C433S mouse (A) The knockin construct, the endogenous LKB1 allele containing exons 1–10 and the target allele with the puromycin cassette (Puro) removed by Flp (flippase) recombinase are depicted. Characterization of the farnesylation-deficient LKB1C433S/C433S mouse (A) The knockin construct, the endogenous LKB1 allele containing exons 1–10 and the target allele with the puromycin cassette (Puro) removed by Flp (flippase) recombinase are depicted. The black/grey rectangles represent exons and the black triangles represent FRT (flippase-recognition target) sites. TK, thymidine kinase. (B) To genotype mice, genomic DNA was PCR-amplified. Sizes are indicated in bp. The breeding strategy employed to generate LKB1C433S/C433S mice is shown with the number and percentage of each genotype obtained indicated. (C) Tissues lysates from wild-type LKB1+/+ and LKB1C433S/C433S mice and MEFs were subjected to immunoblotting using anti-LKB1 and anti-LKB1 farnesylation-specific (LKB1 Farn C433) antibodies. (D) Testis lysates from LKB1+/+ and LKB1C433S/C433S mice were immunoblotted with an anti-LKB1 antibody to detect the LKB1long (LKB1L) and LKB1short (LKB1s) isoforms. Band intensities were quantified using Li-Cor. Results are means±S.E.M. for four mice per genotype. (E) LKB1 was immunoprecipitated (IP) from the liver and muscle (EDL) from wild-type LKB1+/+ (+/+) and LKB1C433S/C433S (C433S/C433S) mice and the in vitro kinase activity towards the LKBtide peptide was measured. Immunoprecipitates were also immunoblotted (IB). Assays were performed in duplicate from tissues taken from six mice per genotype and the results are means±S.E.M. The broken line represents the background activity as measured with pre-immune IgG. (F) As (E), except that the immunoprecipitated LKB1 protein was used to activate a recombinant heterotrimeric AMPK complex (α1β2γ1) and then AMPK kinase activity towards the AMARA peptide was measured. Assays were performed in duplicate from tissues taken from six mice per genotype and the results are means±S.E.M. (G) Liver and MEFs from wild-type (WT) LKB1+/+ (+/+) and LKB1C433S/C433S (KI/KI) mice were submitted to subcellular fractionation. Cytoplasmic, membrane and nuclear fractions were immunoblotted with the indicated antibodies. Blots for two animals out of four per genotype are shown. Vanessa P. Houde et al. Biochem. J. 2014;458:41-56 ©2014 by Portland Press Ltd