List of primary antibodies. Protein target. Supplier and catalogue number. Actin. Sigma-Aldrich. AKT. CST #9272. pAKT S473. CST #4058. S6. CST #2317.
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Oncotarget, Supplementary Materials 2016
The mTOR inhibitor Everolimus synergizes with the PI3K inhibitor GDC0941 to enhance anti-tumor efficacy in uveal melanoma SUPPLEMENTARY DATA SUPPLEMENTARY MATERIALS List of primary antibodies Protein target
Supplier and catalogue number
Actin
Sigma-Aldrich
AKT
CST #9272
pAKT S473
CST #4058
S6
CST #2317
pS6 S235/236
CST #2211
ERK1/2
p44/42 MAPK CST#9102
pERK1/2
Phospho-p44/42 MAPK (T202/Y204) CST #9101
4EBP1
CST #9644
p4EBP1
CST #9459
YAP
CST #4912
pYAP
CST #4911
Oncotarget, Supplementary Materials 2016
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Supplementary Table S1: Genetic profiles of uveal melanoma cell lines screened for combinations Cell line
Supplementary Table S2: Targeted agents used in the combination screen Target
Compound name
PKC
AEB071 (Sotrastaurin)
MEK
GSK1120212 (Trametinib) ; AZD6244 (Selumitinib)
PI3K αβγδ
GDC0941 (Pictilisib)
mTOR
RAD001 (Everolimus)
Dual pan-PI3K/mTOR AKT
BEZ235 KRX0401 (Perifosine)
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Oncotarget, Supplementary Materials 2016
Supplementary Table S3: Gene set enrichment analysis performed on transcriptomic data with David database Below is a screen snap-shot of the results. More details are presented in the corresponding excel file. See Supplementary File 1 See Supplementary File 2
Oncotarget, Supplementary Materials 2016
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Supplementary Table S4: Characteristic of the two PDX models used in the study PDXs
Origin
Histology
GNAQ
GNA11
BAP1
SF3B1
L3
MM66
Liver metastasis
Epithelioid
wt
mut
wt
wt
N
MM52
Liver metastasis
Mixed cells
mut
wt
mut
mut
L
Abbreviations: L3: status of chromosome 3; N = normal, L = loss of heterozygocy -- Mixed cells included epithelioid and spindle cells.
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Oncotarget, Supplementary Materials 2016
Supplementary Figure S1: Excess over Bliss values for the two most synergistic drug combinations. A. PI3Ki + mTORi.
B. PI3Ki + MEKi. PI3Ki = PI3K inhibitor GDC0941; mTORi = mTORC1 inhibitor RAD001 (Everolimus); MEKi = MEK inhibitor AZD6244 (Selumitinib). All compound concentrations are shown in μM.
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Oncotarget, Supplementary Materials 2016
Supplementary Figure S2: Partial matrices obtained in the combination screen for the two most synergistic drug combinations. One matrix is shown per cell line and per combination. Percentages of growth inhibition compared to DMSO-treated control are shown. A color code from blue to red is used to highlight the level of growth inhibition. Names of control lines are written in orange. A. PI3Ki + mTORi. B. PI3Ki + MEKi.
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Oncotarget, Supplementary Materials 2016
Supplementary Figure S3: Full dose matrices for the PI3Ki + mTORi combination. For each cell line, the results of growth inhibition (percentage compared to DMSO-control cells) and the Excess over Bliss values for every tested dose are shown. A color code highlights strong Excess over Bliss values (i.e. synergy) in red.
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Oncotarget, Supplementary Materials 2016
Supplementary Figure S4: Full dose matrices for the PI3Ki + mTORi combination – continued.
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Oncotarget, Supplementary Materials 2016
Supplementary Figure S5: Quantification of cell cycle analyses in Mel202 (A) and MM28 (B) cell lines. Top: quantification of cell populations in sub-G1, cells with 2N, in S or M phases. Bottom: quantification of cell populations in G1, S and G2/M phases. Data are represented as mean ±SEM. Statistical differences between control and treatments in the sub-G1 population was done using two-way ANOVA with with Bonferroni correction. * p