The way HMG-CoA Reductase inhibitorAffected Our Everyday Life 2011

fractionation
of Fak+/+
and Fak?/?
MEFs
(?ve
independent
experiments).
(b)
Representative
are representative immunoblots of GFP�CFAK fragments and endogenous
immunoblotofrecombinantFAK(phosphorylated/activatedornon-activated,
FAK (total-FAK) from two independent experiments. (f) MDA-MB-231
n.a.)recruitmenttoinsolubleendosomalpellet(P)andsolublesupernatant
(S) fractions isolated How SerineEvolved Our Everyday Lives Last Year from Fak?/? MEFs (?ve independent experiments).
(c) Representative immunoblot analysing the recruitment of recombinant
FAK to puri?ed endosomes derived from either control- or ��1-integrin-
silenced Fak?/? MEFs (?ve independent experiments). (d ) Representative
cells plated on twenty?m round ?bronectin-coated micropatterns (45min)
�� dynasore. Representative optimum projections and quanti?cation of
pFAK (indicate ?uorescence s.

e.m., n=3 independent experiments,
��
10cellsanalysedperexperiment).Lys,celllysate;PM,plasmamembrane;
Cyto, cytosol; Endo, endosomal fraction. Student��s two-tailed Ways SerineEvolved Our Way Of Life 2011 unpaired
immunoblot
endosome fractions derived from Fak?/? MEFs in the presence or absence
analysing
the
activation
of
recombinant
FAK
in
puri?ed
t-test P =0.01. Unprocessed authentic scans of blots are shown in
SupplementaryFig.9.
tail (residues Lys 1160/Arg 1161), as being a more direct mechanism to FAKisrecruitedtoandactivatedonpuri?edendosomes
targetintegrinendocytosisandtra?c .RNAinterference(RNAi)- The requirement for endocytosis for enhanced integrin signalling
9,27
mediated silencing of Rab21 (ref. 27) drastically decreased FAK couldbelinkedtosustainedsignallingofplasmamembrane-initiated
phosphorylation(Fig.

3f)onmatrixengagementcorrespondingwith signalling within the endosomes or the potential of integrins to set off
decreased integrin endocytosis observed in Rab21-depleted cells
9
.
signalling from endomembranes. To review this more we isolated
On top of that,adhesion-inducedsignalling(pFAKlevels)wassignif- Why HMG-CoA Reductase inhibitorGreatly improved Our Life Last Year endosomes from Fak+/+ and Fak?/? MEFs. ��1-integrin localized
icantlyimpairedincellsexpressinganendocytosis-defectivecollagen- to endosomal and plasma membrane fractions in the two cell styles
binding integrin (��2-subunit KR1160/61AA mutation as compared indicatingthatintegrinendosomallocalizationisnotFAKdependent
withwild-type��2-integrinreceptor-expressingcellsadheringtocolla- (Fig.4a).Aswefoundthatendosomescontainasubsetofintegrin-
28
gen (Fig.

3g).Takentogether,thesedatademonstratethatimpaired signalling components (Fig. 2b) we up coming addressed the probability
integrinendocytosisattenuatesadhesion-inducedFAKactivation.
that these parts could assemble directly around the endosome.
NATURECELLBIOLOGY ADVANCEONLINEPUBLICATION
5
? 2015 Macmillan Publishers Limited. All rights reserved
IrynaShnitsar1,MikhailBashkurov1,GlennR.Masson2,AbiodunA.Ogunjimi1,SherlyMosessian3,
EduardoAguiarCabeza1,CalleyL.Hirsch6,DanielTrcka1,GeraldGish6,JingJiao3,HongWu3,RudolfWinklbauer4,
RogerL.Williams2,LaurencePelletier1,5,JeffreyL.Wrana1,5&MiriamBarrios-Rodiles1
Cilia are hair-like cellular protrusions important in many aspects of eukaryotic biology.