Hippo pathway

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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.

-

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.

-

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

0.07

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