Hippo pathway - Antibodies Pathway.pdfA6510 GLI2 2736 WB IHC H M R A7159 LATS1 9113 WB IHC H M R...
Transcript of Hippo pathway - Antibodies Pathway.pdfA6510 GLI2 2736 WB IHC H M R A7159 LATS1 9113 WB IHC H M R...
Hippo pathway
ABclonal Antibody(Ⅴ)
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Catalog Name GeneID Reactivity Application
Immunohistochemistry of paraffin-embedded rat kidney using MDM2 antibody at dilution of 1:200 (x400 lens).
A0345
A1351
Western blot analysis of extracts of various cell lines, using CD44 antibody.
A2880
Immunohistochemistry of paraffin-embedded Human cervical cancer tissue using CTGF antibody.
Immunohistochemistry of paraffin-embedded human stomach using MDM2 antibody at dilution of 1:200 (x400 lens).
Hippo pathway list
A7960
Immunofluoresc-ence analysis of A549 cell using ASCC3 antibody. Blue: DAPI for nuclear staining.
A2055
Western blot analysis of extracts of various cell lines, using RUNX1 antibody.
Immunohistochemistry of paraffin-embedded rat kidney using RUNX1 antibody at dilution of 1:100 (x400 lens).
Immunofluorescen ce analysis of MCF-7 cell using RUNX1 antibody. Blue: DAPI for nuclear staining.
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A5761
Immunofluor
-escence analysis of U2OS cell using TP53 antibody.
Immunofluorescence analysis of A549 cell using Tp53 antibody. Blue: DAPI for nuclear staining.
A1675Immunofluorescence analysis of MCF7 cell using PAX3 antibody. Blue: DAPI for nuclear staining.
Hippo pathway list
Western blot analysis of extracts of various cells, using TP53 antibody.
Western blot analysis of extracts of various cells, using TP53 antibody.
A2137
Immunohistochemistry of paraffin-embedded rat kidney using TP63 antibody at dilution of 1:100 (x400 lens).
Immunofluorescence analysis of U2OS cell using TP63 antibody. Blue: DAPI for nuclear staining.
Western blot analysis of extracts of various cell lines,using SMAD1 antibody.
Immunohistochemistry of paraffin-
embedded human stomach cancer using SMAD1 antibody at dilution of 1:200 (400x lens).
-Immunofluorescence analysis of HeLa cell using SMAD1 antibody. Blue: DAPI for nuclear staining
Western blot analysis of extracts of various cell lines, using SMAD4 antibody.
A5657
Immunofluorescence analysis of U20S cell using SMAD4 antibody. Blue: DAPI for nuclear staining.
P0085A
Immunofluorescence analysis of U2OS cell using Phospho-TP53-S9 antibody. Blue: DAPI for nuclear staining.
A1101
A5657
Immunofluorescence analysis of HeLa cell using SMAD4 antibody. Blue: DAPI for nuclear staining.
Immunofluorescence images of SW480 cells showing nuclear and cytoplasmic localization with SMAD4 Antibody 1:200 (top, red).
Western blot analysis of extracts of various cell lines, using STK3 antibody.
Catalog Name GeneID Reactivity Application
A1023 14-3-3 protein beta/alpha 7529 WB IHC IF H M R
A1058 14-3-3 protein epsilon 7531 WB IHC IF H M R
A3043 14-3-3 protein gamma 7532 WB IHC H M R
A6992
A2563 14-3-3 protein theta 10971 WB IHC IF H M R
A7639 14-3-3 protein zeta/delta 7534 WB IHC H
A8075 AMOT 154796 WB IHC H
A1860 AREG 374 WB IHC H M
A7960 ASCC3 10973 WB IF H
A1551 BIRC5 332 WB IHC IF H M R
A1351 CD44 960 WB IHC IF IP H M
A3812 CSNK1D 1453 WB IF IP H M R
A2880 CTGF 1490 WB IHC H M R
A6510 GLI2 2736 WB IHC H M R
A7159 LATS1 9113 WB IHC H M R
A7160 LATS2 26524 WB IHC IF FC H M
A0345 MDM2 4193 WB IHC H M R
A5091 MOBKL1A/MOB1B 92597 WB IHC IP H M R
A4818 MOBKL1B/MOB1A 55233 WB H M
A2456 NF2/Merlin 4771 WB IHC IF H M R
A5761 p53 7157 WB IHC IF H M R
A2137 p63 8626 WB IHC IF H M R
A1675 PAX3 5077 WB IHC IF H M R
AP0083 Phospho-TP53-S15 7157 WB IF H M R
AP0085 Phospho-TP53-S9 7157 WB IF H M
AP0195 Phospho-YWHAZ-S58 7534 WB IHC IF H M R
A0270 PPARG 5468 WB IHC IF H M R
A1551
Western blot analysis of extracts of various cell lines, using BIRC5 antibody.
Immunofluorescence analysis of U2OS cell using BIRC5 antibody. Blue: DAPI for nuclear staining.
Immunohistochemistry of paraffin-embedded human gastric using SMAD3 antibody at dilution of 1:100 (x400 lens).
A7536Immunohistochemistry of paraffin-embedded human kidney using SMAD3 antibody at dilution of 1:100 (x400 lens).
Catalog Name GeneID Reactivity Application
Hippo pathway list Star products
A2055 RUNX1 861 WB IHC ICC IF IP H M R
A1101 SMAD1 4086 WB IHC IF H M R
A0440 SMAD2 4087 WB IHC IF IP H M R
A7536 SMAD3 4088 WB IHC H M R
A5657 SMAD4 4089 WB IHC IF H M R
A6992 STK3/MST2 6788 WB IHC IF H R
A8043 STK4 6789 WB IHC IF IP H M R
A8202 TAZ 25937 WB H M R
A6768 TEAD1 7003 WB IHC H R
A7454 TEAD3 7005 WB IHC H M
A4151 TEAD4 7004 WB IP H M R
A1002 YAP1 10413 WB IHC IF IP FC H M Z
A2195 ZAP70 7535 WB IHC ICC IP H M R
A0594 ZO-2/TJP2 9414 WB IF H M R
A0270
Western blot analysis of extracts of various cell lines, using PPARG antibody.
Immunohistochemistry of
paraffin-embedded H-ovary using PPARG Antibody at Dilution 1:100
-Immunofluore
scence analysis of A549 cell using PPARG antibody. Blue: DAPI for nuclear staining.
-
A3043
Western blot analysis of extracts of various cell lines, using YWHAG antibody.
Immunohistochemistry of
paraffin-embedded Human lung cancer tissue using YWHAG antibody.
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A7639Western blot analysis of extracts of various cell lines, using YWHAZ antibody.
Western blot analysis of extracts of various cell lines, using STK4 antibody.
A8043 A1023
Western blot analysis of extracts of various cell lines, using YWHAB antibody.
A1058
Western blot analysis of extracts of various cell lines, using YWHAE antibody.
A2195
Western blot analysis of extracts of various cell lines, using ZAP70 antibody.
0
0.01
0.02
0.03
0.04
0.05
0.06
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0.08
CTGF GAPDH
Sig
na
l re
lati
ve
to 2
0%
inp
ut
HeLa
TEAD1 IgG
Chromatin immunoprecipitation
analysis extracts of HeLa cell line,
using TEAD1 antibody and rabbit
IgG. The amount of
immunoprecipitated DNA was
checked by quantitative PCR.
Histogram was constructed by the
ratios of the immunoprecipitated
DNA to the input.
Western blot analysis of
extracts of various cell lines,
using TEAD1 antibody.
Chromatin immunoprecipitation
analysis extracts of HeLa cell line,
using TEAD3 antibody and rabbit
IgG. The amount of
immunoprecipitated DNA was
checked by quantitative PCR.
Histogram was constructed by the
ratios of the immunoprecipitated
DNA to the input.
Immunohistochemistry of
paraffin-embedded human
colon using TEAD3 antibody
at dilution of 1:100 (x400
lens).
Because professional so excellence
A6768
A7454
Western blot
analysis of
extracts of
various cell
lines, using NF2
antibody
Immunofluorescence analysis of
HeLa cell using NF2 antibody.
Blue: DAPI for nuclear staining.
A2456Western blot
analysis of
extracts from
HEK 293 cells
and DLD-1
cells using
YAP1 antibody
(1:1000).
Immunofluorescence analysis of
DLD-1 cells showing nuclear and
cytoplasmic localization with YAP1
antibody 1:200 (left,green).
A1002Western blot
analysis of
extracts of
various cell
lines, using
YWHAQ
antibody.
Immunofluorescence analysis of HeLa cell
using YWHAQ antibody. Blue: DAPI for
nuclear staining.
A2563
0
0.005
0.01
0.015
0.02
0.025
CTGFpromoter CTGFpromoter α-satellite
Sig
nal
rela
tive
to 2
0%
inp
ut
HeLa
TEAD3 IgG
A2563
Immunofluorescence analysis of HeLa cell using YWHAQ antibody. Blue: DAPI for nuclear staining.
Western blot analysis of extracts of various cell lines, using YWHAQ antibody.
Western blot analysis of extracts of various cell lines, using AREG antibody.
A1860
P0083AImmunofluorescence analysis of GFP-RNF168 transgenic U2OS cell using
Phospho-TP53-S15 antibody. Green:GFP-RNF168 fusion protein expression for DNA damage marker.Blue: DAPI for nuclear staining. RNF168(GFP) can be used to mark cells damaged by UV-A laser for they always gather around DNA damage region.
The Hippo Signaling PathwayMammals
Hippo signaling is an evolutionarily conserved pathway that controls organ size by regulating cell proliferation,
apoptosis, and stem cell self-renewal. In addition, dysregulation of the Hippo pathway contributes to cancer
development.
The Core of the Mammalian Hippo Pathway
Similar to the invertebrate cascade, a series of phosphorylation events via MST (ortholog of Drosophila Hippo) and Wts
(mammal is LATS1 / 2) ultimately leads to the phosphorylation of two co-activators: YAP and TAZ. Once
phosphorylated, the protein kinase is activated, resulting in a series of changes in downstream processes.
Activation of the Mammalian Hippo Pathway
Yorkie (Yki) is a transcriptional co-activator. As its name, Yki cannot be directly combined with DNA, but it needs to
combine with the transcription factor Scalloped (Sd) to form Yki-Sd complex, which plays a role in transcription
DsFt
Ds
Ft
Ex
kbr
Mer
SavHpo
MtsWts
YkiTF
Known target genes Novel target genes Negative feedback loops
cycE,e2f1,wg,dally,vein,diap1
dpp,wnt6,spz3,pvf2,nplp1,path,CG1607,tsp,timp,foxo,CG4096,InR
ex,wts,ft,ds,kbr
The Hippo Signaling PathwayIkmi A, et al.Molecular evolution of the Yap/Yorkie proto-oncogeneand
elucidation of its core transcriptional program.Mol Biol Evol. 2014 Jun;
31(6):1375-90.
in Drosophila.
regulation. Yki-Sd transcription complex can be
regulated multiple genes related to cell proliferation
and transcription regulation, such as E cyclin, etc..
When the organ reaches a certain size, the pathway
is activated and phosphorylated (activated) Warts
can be phosphorylated by Yki, and the 168th serine
i s ac t i va ted . Yk i b inds to 14-3-3 a f te r
phosphorylation (inactivation). This process allows
the Yki to reside in the cytoplasm, which cannot form
a transcriptional regulatory complex with Sd, and
inhibit transcription of downstream target genes.
In the nucleus, both YAP and TAZ can bind to
TEAD1/4 and activate the transcription of genes
required to promote cell growth and inhibit apoptosis
(CTGF, AREG, BIRC5-2, FGF, and GLI-2). Upon
phosphorylation by the SRC/ABL kinases, YAP and
TAZ acquire the capacity to bind p73 and activate the
transcription of proapoptotic genes. Numerous
transcription factors are described as YAP/TAZ
binding partners, but their functional significance is
still unclear.
The table of proteins:
Dachsous(Ds)
Drosophila FunctionHuman
atypical cadherin / may be a ligand of Fat receptor DCHS1, DCHS2
Fat (Ft)FAT1, FAT2, FAT3,FAT4(FATJ)
atypical cadherin / may be Hippo signaling receptors
Dachs (Dachs) one of the myosin / binding Wts and promoting its degradation
Kibra (Kibra) containing WW domains / combined with Mer and Ex, regulating upstream signal in the pathwayWWC1
Merlin (Mer) containing FERM domains / bind to Kibra and Ex to regulate upstream signal in the pathway NF2
Hippo (Hpo) Sterile-20-like kinase / both phosphorylating and activating the Wts MST1, MST2
Salvador (Sav) containing WW domains which can play a role of scaffold proteinsWw45 (SAV1)
Warts (Wts) DBF-2 related kinase / phosphorylating Yki to inactivate it LATS1, LATS2
Mob as tumor suppressor (Mats)
kinase combine with Wts / The catalytic activity of Wts can be promoted by thecombination.
MOBKL1A, MOBKL1B
Yorkie (Yki) transcription co-activators / combining with transcription factor Sd in the state ofnonphosphorylated activation and activating the transcription of downstream target genes
YAP, TAZ
transcription factors combined with Yki /transcription of target genesScalloped (Sd) TEAD1, TEAD2, TEAD3, TEAD4
Expanded (Ex) containing FERM domains / binding to Kibra and Mer to regulate upstream signal in the pathway FRMD6/Willin