The LuTHy procedure The LuTHy procedure Schematic representation of the workflow of the LuTHy method. Expression vectors encoding NL and PA‐mCit‐tagged.

Slides:

Advertisements

Similar presentations

Tuning the response of the DPI

Advertisements

Nur77 interacts with COUP‐TF in yeast.

Abundance of proteins matching selected subcellular locations and functions in CaCo‐2 cells. Abundance of proteins matching selected subcellular locations.

JNK is activated by Wnt5a.

CNA‐driven network of glioblastoma.

Epistatic interactions and gene expression variation.

Inhibition of SERCA by cyclopiazonic acid normalises both the early decrease and late increase in cytosolic calcium and enhances the viability in differentiated.

Do you know population SD? Use Z Test Are there only 2 groups to

JNK, FAK, Src, and BET inhibitors overcome the NGFRHigh drug‐resistant state in additional BRAFV600E/D melanoma lines JNK, FAK, Src, and BET inhibitors.

Validation of the CCE associations by mass spectrometry.

Identification of progenitor states in human cord blood hematopoiesis

Clustering of CD34+ single cells

Differential accumulation of substrates in vivo

Comparative analysis of RNA and protein profiles.

Disease‐causing mutations influence the association of CSPα with partner proteins Disease‐causing mutations influence the association of CSPα with partner.

Large‐scale image‐based CRISPR‐Cas9 gene perturbation profiling

Correlating protein to mRNA and proteins per mRNA ratios

TGF‐β1 and Sema3A are downregulated in vivo by increasing VEGF doses Muscles were harvested 7 days after implantation of V Low, V Med, and V High clones.

Enhancement of basal mitophagy by USP30 depletion is dependent on PINK1 Enhancement of basal mitophagy by USP30 depletion is dependent on PINK1 Representative.

Network derived from large‐scale fractionation predicts 48 protein complexes and communities Network derived from large‐scale fractionation predicts 48.

Characterization of promoters relative to pAGA1

No neurodegenerative phenotype observed in single Aph1a and Aph1bc cKO Cre+ mice No neurodegenerative phenotype observed in single Aph1a and Aph1bc cKO.

USP30 restricts basal pexophagy

Heat maps showing global relative growth phenotype and comparison between measured and predicted values. Heat maps showing global relative growth phenotype.

WDR11 cooperates with Hh pathway to regulate gene expression

Cycling resident cardiomyocytes in the peri‐infarct area. A–C

PERK activator treatment protects neurite network

Networks of ERK substrates.

Effect of Zc3h10 overexpression in myoblasts and myotubes

Effects of missense mutations and small molecules on PPI detection with LuTHy Effects of missense mutations and small molecules on PPI detection with LuTHy.

Collective shifts of abundance for nuclear and extracellular proteins in colorectal cancer Collective shifts of abundance for nuclear and extracellular.

Effect of the loss of Kar4 on the induction of various promoters

Systematic analysis of interacting proteins with known binding affinities Systematic analysis of interacting proteins with known binding affinities Selection.

Analysis of the phosphomimetic ProP‐PD selection results

Akt inhibition with AZD5363 does not enhance the radiosensitivity of tumour cells in vitro Akt inhibition with AZD5363 does not enhance the radiosensitivity.

ER stress response and susceptibility to apoptosis are regulated by TFEB and TFE3 ER stress response and susceptibility to apoptosis are regulated by TFEB.

IL‐23‐induced psoriasis‐like skin disease is ameliorated in IL‐23−/− mice IL‐23‐induced psoriasis‐like skin disease is ameliorated in IL‐23−/− mice ARepresentative.

Validation of ADORA2A and GPR37 interaction in HEK‐293 cells and native tissue Validation of ADORA2A and GPR37 interaction in HEK‐293 cells and native.

Francis Crick's central dogma of biology revolves around the transcription of mRNA from DNA, the translation of proteins from mRNA, and the degradation.

Schematic representation of LuTHy

Notch activation in Notch1 positive feedback knock‐out intestine organoids Notch activation in Notch1 positive feedback knock‐out intestine organoids Intestine.

QRT–PCR analysis of tissue‐specific candidate gene expression in response to reduced IIS qRT–PCR analysis of tissue‐specific candidate gene expression.

Evaluation of assay performance with positive and negative binary reference sets Evaluation of assay performance with positive and negative binary reference.

PGC‐1α‐independent regulation of the slow‐twitch muscle fiber program by miR‐499 PGC‐1α‐independent regulation of the slow‐twitch muscle fiber program.

UUKV NSs protein inhibits IFN-β induction at the level of MAVS, whereas HRTV NSs antagonizes at the level of TBK1. UUKV NSs protein inhibits IFN-β induction.

TGFβ signaling activity in smooth muscle cells in human left main coronary arteries with various degrees of atherosclerosis TGFβ signaling activity in.

Comparison of proteomics and RNA‐Seq data.

EnAd‐EpCAMBiTE shows reproducible activity within an expanded cohort of patient malignant peritoneal and pleural exudates EnAd‐EpCAMBiTE shows reproducible.

DOK7 gene therapy suppresses motor nerve terminal degeneration at the NMJ in ALS mice DOK7 gene therapy suppresses motor nerve terminal degeneration at.

Simulation showing that the cell length variability of the entire population can mask abnormal cell length variability at a specific cell cycle period.

miR‐34a‐5p acts in PDGFRα+ cells to block lung alveolarization

Protein interactions at the nuclear pore.

Melting behavior of protein complexes

Nedd4 proteins. Nedd4 proteins. (A) Schematic representation of rNedd4‐1, hNedd4‐1 and hNedd4‐2 (not to scale), with % amino‐acid identity between them.

Spatial regulation of Aux/IAA‐ARF signalling in the inflorescence.

Global and focused views of human interaction map.

IIS‐dfoxo‐dependent regulation of proteasome activity in the gut enhances proteome maintenance IIS‐dfoxo‐dependent regulation of proteasome activity in.

The CHCH domain is necessary for mitochondrial import of CHCHD10

Interaction of Gpr1/Gpa2 and Sch9.

Tissue‐specific differences in the levels of CoQ9, CoQ10, and DMQ9, and in the DMQ9/CoQ9 ratio after β‐RA treatment in Coq9R239X mice Tissue‐specific differences.

Sox2 is essential for SLG regeneration following radiation injury

Shape‐sensitivity profiles inferred using shape projection

AS aggregates interact with SERCA in SH‐SY5Y cells

Systematic analysis of protein compensation in a heterozygous strains.

Proteomic analysis of human transcription factors Disease correlation of 19 TFs and 4 well‐studied FOX family members, based on their GO annotations. Proteomic.

Symptomatic rescue of double‐transgenic mice without decreased protein aggregation The expression of exogenous HMGB1‐FLAG (detected with an anti‐FLAG antibody)

Averages from seven separate experiments of primary myoblast cultures grown in HS-2; HS-2+GAL-M; HS-2+COS-1; FCS-20; FCS-20+GAL-M; or FCS-20+COS-1. Averages.

Leptin and amylin signaling.

Histone modification status at the Tβ4 promoter and at the protein level in the HuH7 sublines. Histone modification status at the Tβ4 promoter and at the.

Association of TCTP with Pim-3 in human pancreatic cancer cell lines.

Presentation transcript:

The LuTHy procedure The LuTHy procedure Schematic representation of the workflow of the LuTHy method. Expression vectors encoding NL and PA‐mCit‐tagged hybrid proteins are cloned and co‐transfected into mammalian cells. Binary interactions are detected with a double readout; first, in vivo with in‐cell bioluminescence resonance energy transfer (BRET) and second, ex vivo with a luminescence‐based co‐precipitation (LuC).Schematic representation of control proteins and fusion proteins of interest for investigating the interaction between PA‐mCit‐BAD and NL‐BCL2L1 in proof‐of‐principle LuTHy experiments.Calculated BRET ratios for the indicated protein pairs. The positive control protein PA‐mCit‐NL and the interacting fusion proteins NL‐BCL2L1 and PA‐mCit‐BAD show high BRET ratios.Calculated LuC ratios for the indicated protein pairs. The positive control proteins PA‐NL and PA‐mCit‐NL and the interacting proteins NL‐BCL2L1 and PA‐mCit‐BAD show high LuC ratios.Data information: Data are representative of more than three independent experiments. All values are mean ± s.d. Significance was calculated by one‐way ANOVA followed by Dunnett's multiple comparisons post hoc test; ***P < 0.001. Philipp Trepte et al. Mol Syst Biol 2018;14:e8071 © as stated in the article, figure or figure legend