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Table 2.

Clinically used anticancer agents and their correlations with SLFN11 transcript

Our statistical observations were surprising because the Schlafen family of genes has been studied mainly in the context of immune processes, mouse embryonic lethality, meiotic drive, and orthopoxvirus virulence ( 16 ).

To test the causal relationship between SLFN11 expression and sensitivity to DDAs, we transfected SLFN11-targeting siRNAs in two different human cancer cell lines, DU-145 (prostate cancer) and HOP-62 (non-small-cell lung cancer). Both express high SLFN11 levels and are highly sensitive to Top1 inhibitors ( Easy Spirit Tunein B1qIq5qHr
). In both cell lines, siRNA-mediated silencing proved to be highly efficient and was sustained for 3 d or more ( Fig. 2 A , Lower Right , and Fig. S6 A ). SLFN11-silenced cells exhibited at least a fivefold reduced sensitivity to CPT, etoposide, and cisplatin compared with mock-silenced cells ( P ≤ 0.01, Fig. 2 A , Upper , and Fig. S6 A ). These results demonstrate that SLFN11 confers sensitivity to Top1 inhibitors, Top2 inhibitors, and alkylating agents. In agreement with our bioinformatic predictions, SLFN11 silencing did not affect the cellular response to either the tubulin poison paclitaxel or the broad-spectrum protein kinase inhibitor and apoptosis inducer staurosporine ( 17 ) ( Fig. 2 A , Lower, and Fig. S6 A ).

Fig. 2.

Silencing SLFN11 significantly reduces sensitivity to different classes of DNA-damaging agents in cells expressing high endogenous SLFN11 levels. () Cytotoxicity curves of the prostate cancer cell line DU-145 (high SLFN11 expresser) transfected with nontargeting (ctrl) or SLFN11-targeting siRNAs and treated for 72 h with CPT, etoposide, cisplatin, taxol, or staurosporine (STS). Mean values ± SD are shown (one representative experiment performed in triplicate). () Western blot showing SLFN11 knockdown 3 and 6 d after transfection with SLFN11-targeting siRNAs. () () Representative image of a clonogenic assay (100 nM CPT for 1 d). () Number of colonies formed after 24 h treatment with CPT followed by a 15-d release (average of three independent experiments). () Cytotoxicity curves of the breast cancer cell line MDA-MB-231 (low SLFN11 expresser) transfected with nontargeting (ctrl) or SLFN11-targeting siRNAs and treated for 72 h with CPT, etoposide, and cisplatin. Mean values ± SD are shown (one representative experiment performed in triplicate). Whiskers in all charts represent SDs (when not visible they were within the symbol size). () FACS analysis showing sustained cell cycle progression and lack of apoptosis in SLFN11 knockdown DU-145 cells treated with 100 nM CPT for 16 h.

The influence of SLFN11 on long-term cell survival after CPT treatment was further tested by clonogenic assays. Three days after SLFN11 knockdown, DU-145 cells were treated with 100 nM CPT for 24 h and then released in fresh medium and grown for 15 d to allow colony formation. SLFN11-silenced cells were able to form multiple growing colonies (representative image in Nike Womens Air Max Thea LX Lifestyle Shoes IRhi1SGzd
). Similar results were obtained after treatment with different concentrations of CPT (20–500 nM for 24 h) or after a 1-h incubation with 1 μM CPT followed by a 15-d drug-free growth ( P < 0.01; Fig. 2 B , Right , and Fig. S6 C ).

Horse Adoption

Resist Conquistadors

Garra Charrua

The Last Charruas

Blundstone BL1459 nUCmPF0

Moundbuilders Legacy

Massacre of the Ani-Kutani

Red Chiefs

Cherokee Adaptability

Ghigau

Mountainous Isolation

Cherokee Emperor

Chiba

Taira Lineage

Dream of Masakado

Coalition With Kamakura Kubo

Sakura Kadan

Katori Jingu

Irrigation of Southern Kanto

Naval Reform

McKaria Suede Block Heel Dress Sandals 1o5qg3ZP

Spartans of the Lower Mississippi

Nighttime Raids

Unconquered and Unconquerable

Slave Raids

Lessons of Ackia

Swan Feathers Mantle

Dolce Vita Barkley Woven Cork Wedge Sandal cYJhm7rUk

Inter-Valley Irrigation

Ciudadelas

Split Inheritance

Watchfulness of the Moon

Artisan Relocations

Specialized Economy

The Mit'a

Anne Klein Findaway Tiojwktg

Ichiryo Gusoku

Seven Clans of Tosa

Unification of Shikoku

100 Article Code

Expand the Navy

God of Arts

Support of the Temples

GH Bass amp; Co Margie 20 M91IaM4E

Horsemen of the Steppe

Merchants Of Genoa

Circassian Grain Trade

Franciscan Missionaries

Adyghe Beauty

Steppe Diplomats

Religious Flexibility

Clanricarde

The Fourteen Tribes

By Fire and Sword

People of the Sea

Irish Hobbies

To Hell or to Connacht

Catholic Holdouts

The Lord Presidency

Cleves

Walled Cities

Cleves and Mark

Avid Reformers

Strategic Marriages

United Duchies

Educated Duisberg

Militarize Schwanenburg

Corfu

Frankokratia

Castles of the Angels

Settling the Islands

Gate to the Adriatic

Naval Expansion

Entrepôt of the Eastern Mediterranean

Ionian Academy

Ignite Limitless Metallic Suede Sneakers v8jHM

Stannary Parliaments

Cornish Rebels

Somali sailors

East African Gold Trade

City of Mogadishu

Mogadishan Currency

Horn of Africa

Toob Benadir

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First Dismounting

Razesi Peasants

Upper Boyars

Hospodar of Moldavia

Metropolis of Moldavia

Vlachs

Moldavian Trade

SKECHERS Work Cozard 73ow8Rf

Legacy Of Diocleia

Metropolitanate of Montenegro

Clan Society

Balkan Gusars

The Call Of The Sea

Tax Reform

Cojstvo I Junastvo

Easy Street Kelsa Tall Boot 95oIyYg6

Margraviate of Moravia

Religious Sanctuary

Cities of Moravia

Land of Vineyards

German Quarters

Jesuit University

Legacy of Great Moravia

Descendants of Oe Hiromoto

Leader of Kokujin Ikki

Diplomatic Intrigues

Three Arrows

Hiroshima

Itsukushima

Hagi Ware

Mossi / Yatenga

Riale and Yennenga

Cavalry Raids

Nyonnyonse Vassals

Nakomse

Land of the Ancestors

Honoring the Masks

Friday Court

Elegant Footwear Dollie Platform Ankle Strap Sandal RssTh

An Indian Padishah

The Jagir and Mansabdari Systems

Control over the Clergy

Ibadat Khana - House of Worship

Zabt System

Mughal Architecture

Fatawa-i-Alamgiri

Muisca

Zipa and Zaque

El Dorado

Gueches Wrestling

The Tale of Bochica

Emeralds Barter

Terrace Farming

Consuetudinary Law

Munster

North Sea Fishermen

The Brave and Faithful

Hill Raiders

Spanish Ties

Fairer Shores

Pirate Haven

Enemies Within

Münster

Great Procession

Erbmänner

Founding of Monasteries

Cities of peace

Soldiers of the Imperial Armies

Bommen Berend

Support the Schools

Mephisto Josia rq1Jp

Gatherers of Tribute

Legacy of Dmitriy Donskoi

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