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ES cells could be maintained in culture but were generally slower growing than Atrx+ ES clones, and appeared to undergo higher rates of spontaneous differentiation. We investigated directly the effect of Atrx on ES cell growth by comparing Atrx+ and Atrxnull ES cell clones in competition cultures. Equal numbers of Atrx+ (bearing either an Atrx WT or an Atrx flox allele) and Atrxnull (bearing an Atrx Δ18Δneo allele) ES cells were inoculated into cultures and the mixed cultures were passaged (1:3 split) every 2 d for 8–10 d. The relative abundance of the different alleles in the culture at each time point was analysed by Southern blotting (Figure 3A). The clone containing the Atrx Δ18Δneo allele was rapidly outgrown by both Atrx WT ES cells and cells bearing the Atrx flox allele. In a control competition experiment between different clones bearing functional Atrx alleles (Atrx WT and Atrx flox), both clones continued to be equally represented after 8 d of cocultivation. Thus, although Atrxnull ES cells could be recovered and maintained in culture by a conditional targeting approach, these cocultivation experiments suggested that the absence of Atrx does negatively impact upon normal ES cell growth."}

    craft-ca-core-ex-dev

    {"project":"craft-ca-core-ex-dev","denotations":[{"id":"T2328","span":{"begin":0,"end":4},"obj":"PR_EXT:000004503"},{"id":"T2329","span":{"begin":4,"end":8},"obj":"SO_EXT:sequence_nullness"},{"id":"T2330","span":{"begin":9,"end":17},"obj":"CL:0002322"},{"id":"T2331","span":{"begin":12,"end":17},"obj":"CL_GO_EXT:cell"},{"id":"T2332","span":{"begin":75,"end":82},"obj":"GO:0040007"},{"id":"T2333","span":{"begin":88,"end":92},"obj":"PR_EXT:000004503"},{"id":"T2334","span":{"begin":94,"end":96},"obj":"CL:0002322"},{"id":"T2335","span":{"begin":97,"end":103},"obj":"SO_EXT:sequence_cloned_entity"},{"id":"T2336","span":{"begin":157,"end":172},"obj":"GO_RO_EXT:developmental_differentiation_process"},{"id":"T2337","span":{"begin":213,"end":217},"obj":"PR_EXT:000004503"},{"id":"T2338","span":{"begin":221,"end":228},"obj":"CL:0002322"},{"id":"T2339","span":{"begin":224,"end":228},"obj":"CL_GO_EXT:cell"},{"id":"T2340","span":{"begin":229,"end":235},"obj":"GO_EXT:biological_growth_entity_or_process"},{"id":"T2341","span":{"begin":249,"end":253},"obj":"PR_EXT:000004503"},{"id":"T2342","span":{"begin":259,"end":263},"obj":"PR_EXT:000004503"},{"id":"T2343","span":{"begin":263,"end":267},"obj":"SO_EXT:sequence_nullness"},{"id":"T2344","span":{"begin":268,"end":275},"obj":"CL:0002322"},{"id":"T2345","span":{"begin":271,"end":275},"obj":"CL_GO_EXT:cell"},{"id":"T2346","span":{"begin":276,"end":282},"obj":"SO_EXT:sequence_cloned_entity"},{"id":"T2347","span":{"begin":325,"end":329},"obj":"PR_EXT:000004503"},{"id":"T2348","span":{"begin":350,"end":354},"obj":"PR_EXT:000004503"},{"id":"T2349","span":{"begin":355,"end":357},"obj":"SO_EXT:wild_type_entity_or_quality"},{"id":"T2350","span":{"begin":364,"end":368},"obj":"PR_EXT:000004503"},{"id":"T2351","span":{"begin":369,"end":373},"obj":"SO:0000359"},{"id":"T2352","span":{"begin":374,"end":380},"obj":"SO_EXT:0001023"},{"id":"T2353","span":{"begin":386,"end":390},"obj":"PR_EXT:000004503"},{"id":"T2354","span":{"begin":390,"end":394},"obj":"SO_EXT:sequence_nullness"},{"id":"T2355","span":{"begin":407,"end":411},"obj":"PR_EXT:000004503"},{"id":"T2356","span":{"begin":420,"end":426},"obj":"SO_EXT:0001023"},{"id":"T2357","span":{"begin":428,"end":436},"obj":"CL:0002322"},{"id":"T2358","span":{"begin":431,"end":436},"obj":"CL_GO_EXT:cell"},{"id":"T2359","span":{"begin":578,"end":585},"obj":"SO_EXT:0001023"},{"id":"T2360","span":{"begin":671,"end":676},"obj":"SO_EXT:sequence_cloned_entity"},{"id":"T2361","span":{"begin":692,"end":696},"obj":"PR_EXT:000004503"},{"id":"T2362","span":{"begin":705,"end":711},"obj":"SO_EXT:0001023"},{"id":"T2363","span":{"begin":741,"end":745},"obj":"PR_EXT:000004503"},{"id":"T2364","span":{"begin":746,"end":748},"obj":"SO_EXT:wild_type_entity_or_quality"},{"id":"T2365","span":{"begin":749,"end":757},"obj":"CL:0002322"},{"id":"T2366","span":{"begin":752,"end":757},"obj":"CL_GO_EXT:cell"},{"id":"T2367","span":{"begin":762,"end":767},"obj":"CL_GO_EXT:cell"},{"id":"T2368","span":{"begin":780,"end":784},"obj":"PR_EXT:000004503"},{"id":"T2369","span":{"begin":785,"end":789},"obj":"SO:0000359"},{"id":"T2370","span":{"begin":790,"end":796},"obj":"SO_EXT:0001023"},{"id":"T2371","span":{"begin":852,"end":858},"obj":"SO_EXT:sequence_cloned_entity"},{"id":"T2372","span":{"begin":878,"end":882},"obj":"PR_EXT:000004503"},{"id":"T2373","span":{"begin":883,"end":890},"obj":"SO_EXT:0001023"},{"id":"T2374","span":{"begin":892,"end":896},"obj":"PR_EXT:000004503"},{"id":"T2375","span":{"begin":897,"end":899},"obj":"SO_EXT:wild_type_entity_or_quality"},{"id":"T2376","span":{"begin":904,"end":908},"obj":"PR_EXT:000004503"},{"id":"T2377","span":{"begin":909,"end":913},"obj":"SO:0000359"},{"id":"T2378","span":{"begin":921,"end":927},"obj":"SO_EXT:sequence_cloned_entity"},{"id":"T2379","span":{"begin":1007,"end":1011},"obj":"PR_EXT:000004503"},{"id":"T2380","span":{"begin":1011,"end":1015},"obj":"SO_EXT:sequence_nullness"},{"id":"T2381","span":{"begin":1016,"end":1024},"obj":"CL:0002322"},{"id":"T2382","span":{"begin":1019,"end":1024},"obj":"CL_GO_EXT:cell"},{"id":"T2383","span":{"begin":1169,"end":1173},"obj":"PR_EXT:000004503"},{"id":"T2384","span":{"begin":1209,"end":1216},"obj":"CL:0002322"},{"id":"T2385","span":{"begin":1212,"end":1216},"obj":"CL_GO_EXT:cell"},{"id":"T2386","span":{"begin":1217,"end":1223},"obj":"GO_EXT:biological_growth_entity_or_process"}],"text":"Atrxnull ES cells could be maintained in culture but were generally slower growing than Atrx+ ES clones, and appeared to undergo higher rates of spontaneous differentiation. We investigated directly the effect of Atrx on ES cell growth by comparing Atrx+ and Atrxnull ES cell clones in competition cultures. Equal numbers of Atrx+ (bearing either an Atrx WT or an Atrx flox allele) and Atrxnull (bearing an Atrx Δ18Δneo allele) ES cells were inoculated into cultures and the mixed cultures were passaged (1:3 split) every 2 d for 8–10 d. The relative abundance of the different alleles in the culture at each time point was analysed by Southern blotting (Figure 3A). The clone containing the Atrx Δ18Δneo allele was rapidly outgrown by both Atrx WT ES cells and cells bearing the Atrx flox allele. In a control competition experiment between different clones bearing functional Atrx alleles (Atrx WT and Atrx flox), both clones continued to be equally represented after 8 d of cocultivation. Thus, although Atrxnull ES cells could be recovered and maintained in culture by a conditional targeting approach, these cocultivation experiments suggested that the absence of Atrx does negatively impact upon normal ES cell growth."}

    craft-ca-core-dev

    {"project":"craft-ca-core-dev","denotations":[{"id":"T2215","span":{"begin":0,"end":4},"obj":"PR:000004503"},{"id":"T2216","span":{"begin":9,"end":17},"obj":"CL:0002322"},{"id":"T2217","span":{"begin":75,"end":82},"obj":"GO:0040007"},{"id":"T2218","span":{"begin":88,"end":92},"obj":"PR:000004503"},{"id":"T2219","span":{"begin":94,"end":96},"obj":"CL:0002322"},{"id":"T2220","span":{"begin":213,"end":217},"obj":"PR:000004503"},{"id":"T2221","span":{"begin":221,"end":228},"obj":"CL:0002322"},{"id":"T2222","span":{"begin":249,"end":253},"obj":"PR:000004503"},{"id":"T2223","span":{"begin":259,"end":263},"obj":"PR:000004503"},{"id":"T2224","span":{"begin":268,"end":275},"obj":"CL:0002322"},{"id":"T2225","span":{"begin":325,"end":329},"obj":"PR:000004503"},{"id":"T2226","span":{"begin":350,"end":354},"obj":"PR:000004503"},{"id":"T2227","span":{"begin":364,"end":368},"obj":"PR:000004503"},{"id":"T2228","span":{"begin":369,"end":373},"obj":"SO:0000359"},{"id":"T2229","span":{"begin":374,"end":380},"obj":"SO:0001023"},{"id":"T2230","span":{"begin":386,"end":390},"obj":"PR:000004503"},{"id":"T2231","span":{"begin":407,"end":411},"obj":"PR:000004503"},{"id":"T2232","span":{"begin":420,"end":426},"obj":"SO:0001023"},{"id":"T2233","span":{"begin":428,"end":436},"obj":"CL:0002322"},{"id":"T2234","span":{"begin":578,"end":585},"obj":"SO:0001023"},{"id":"T2235","span":{"begin":692,"end":696},"obj":"PR:000004503"},{"id":"T2236","span":{"begin":705,"end":711},"obj":"SO:0001023"},{"id":"T2237","span":{"begin":741,"end":745},"obj":"PR:000004503"},{"id":"T2238","span":{"begin":749,"end":757},"obj":"CL:0002322"},{"id":"T2239","span":{"begin":780,"end":784},"obj":"PR:000004503"},{"id":"T2240","span":{"begin":785,"end":789},"obj":"SO:0000359"},{"id":"T2241","span":{"begin":790,"end":796},"obj":"SO:0001023"},{"id":"T2242","span":{"begin":878,"end":882},"obj":"PR:000004503"},{"id":"T2243","span":{"begin":883,"end":890},"obj":"SO:0001023"},{"id":"T2244","span":{"begin":892,"end":896},"obj":"PR:000004503"},{"id":"T2245","span":{"begin":904,"end":908},"obj":"PR:000004503"},{"id":"T2246","span":{"begin":909,"end":913},"obj":"SO:0000359"},{"id":"T2247","span":{"begin":1007,"end":1011},"obj":"PR:000004503"},{"id":"T2248","span":{"begin":1016,"end":1024},"obj":"CL:0002322"},{"id":"T2249","span":{"begin":1169,"end":1173},"obj":"PR:000004503"},{"id":"T2250","span":{"begin":1209,"end":1216},"obj":"CL:0002322"}],"text":"Atrxnull ES cells could be maintained in culture but were generally slower growing than Atrx+ ES clones, and appeared to undergo higher rates of spontaneous differentiation. We investigated directly the effect of Atrx on ES cell growth by comparing Atrx+ and Atrxnull ES cell clones in competition cultures. Equal numbers of Atrx+ (bearing either an Atrx WT or an Atrx flox allele) and Atrxnull (bearing an Atrx Δ18Δneo allele) ES cells were inoculated into cultures and the mixed cultures were passaged (1:3 split) every 2 d for 8–10 d. The relative abundance of the different alleles in the culture at each time point was analysed by Southern blotting (Figure 3A). The clone containing the Atrx Δ18Δneo allele was rapidly outgrown by both Atrx WT ES cells and cells bearing the Atrx flox allele. In a control competition experiment between different clones bearing functional Atrx alleles (Atrx WT and Atrx flox), both clones continued to be equally represented after 8 d of cocultivation. Thus, although Atrxnull ES cells could be recovered and maintained in culture by a conditional targeting approach, these cocultivation experiments suggested that the absence of Atrx does negatively impact upon normal ES cell growth."}