Six Profiting Tips For Temozolomide Which Usually never Fails

opy detected spherical, membrane-
mutations)intumourcellswereassessed(Fig.2f).ComparedwithCAF encapsulated particles between 30 and 100nm, typical of exosome
co-culture,astrocyteco-cultureinhibitedluciferaseactivityofwild-type vesicles,inastrocyte-conditionedmedia (Fig.3b).Additionally,the
22
PTEN39-UTR,whichwasrescuedbythemiR-19abindingsitemuta- Five Sensational Ideas For Temozolomide That Usually never Fails astrocyte-conditioned media contained significantly more CD631,
tion(position1),butnotbyothermutations,indicatingthemajorrole CD811 andTSG1011 exosomes
thantheCAF-conditionedmedia
22
of miR-19a in astrocyte-mediated PTEN mRNA downregulation in (Fig.3candExtendedDataFig.4e,f).Moreover,theexosomesfrom
tumour cells. Furthermore, PTEN mRNA (Fig. 2g and Extended astrocytescontained3.5-foldhigherlevelsofmiR-19athanthosefrom
DataFig.

3g)andPTENprotein(Fig.2handExtendedDataFig.3h) CAFs(ExtendedDataFig.4g).Addingexosomespurifiedfromcondi-
were not downregulated in tumour cells Two Successful Suggestions For Loratadine Which Practically never Fails co-cultured with primary tionedmediaofCy3-miR-19a-transfectedastrocytesledtomiR-19a
astrocytes from Mirc1tm1.1Tyj/J mice in which PTEN-targeting transfer into cultured tumour cells (Fig. 3d). Furthermore, treating
miRNAsweredepleted(ExtendedDataFig.3i).
tumourcellsdirectlywithastrocyte-derivedexosomesledtoadose-
After co-culture with Cy3-labelled miR-19a-transfected primary dependentincreaseofmiR-19aandasubsequentdecreaseofPTEN
astrocytes,wedetectedsignificantlymoreCy31epithelialcelladhesion mRNA in tumour cells (Fig. 3e). To determine whether astrocyte-
molecule(EpCAM)-positivetumourcellsovertimethanunderCAF released exosomes are required for miR-19a transfer, we blocked
co-culture(Fig.

3aandExtendedDataFig.4a),suggestingthatmiR-19a astrocyteexosomesecretionbytreatingastrocyteswitheitheraninhib-
isintercellularlytransferredfromastrocytestotumourcells.miRNAs 15 Success Suggestions For Loratadine That Usually never Fails itorofexosomerelease,dimethylamiloride(DMA),orashortinter-
aretransferablebetweenneighbouringcellsthroughgapjunctionsor fering RNA (siRNA) targeting Rab27a, a mediator of exosome
19,20
smallvesicles .Treatingtumourcellswithagapjunctionchannel
secretion (Extended Data Fig. 5a�Cc). Both exosome blockades
23
a
b
d
h
i
HCC1954
1. Intracranial injection
of lentiviral shRNAs
shControl
M
shRab27a/b
1954P + exosome from Cy3-miR-19a astrocyte
50 ��m
M
Astrocyte 12 h
HCC1954
B
B
shControl
shRab27a/b
50 ��m
50 ��m
M
M
50 nm
astrocyte
3.

16
2. Intracranial injection
of B16BL6
100
80
60
40
20
0
B
B
60 h
j
k
c
CAF
1.00
1. Intracranial injection
of lentiviral shRab27a/b
3
2
1
P < 0.05
CD63
CD81
�C103
0
103 104 105 �C103
Cy3
0
103 104 105
P = 0.0157
Cy3
n = 7
2. Intracranial
injection
2. Intracranial
injection
0.5
0.0
1.00
1.00
4.09
+Supernatant
+Exosome
Negative control
Positive control
0
TSG101
Cy3-miR-19
CAF
Cy3-miR-19
astrocyte
�C0.5
�C1.0
�C1.5
�C2.0
B16BL6
+ vehicle
B16BL6
6.78
HCC1954 co-culture
+ astrocyte-derived
exosome
e
f
g
miR-19a
PTEN
miR-19a
PTEN
miR-19a
PTEN
m
2.5
2.0
1.5
1.0
0.5
0.0
P = 0.0163
shControl shRab27a/b
P = 0.0042
P = 0.0002
1.5
1.0
0.5
0.0
1.5
P = 0.036 P = 0.012
P = 0.0023
B16BL6 + exosome
1.0
0.5
0.0
l
B16BL6
P = 0.0091
n = 8
2.0
1.0
50 ��m
M
M
B
B
0.0
�C1.0
�C2.0
M
M
Vehicle