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mL FGF-2刺激细胞不同时间并裂解。使用对磷酸化的p44 \u002F 42促分裂原活化蛋白激酶（Thr202 \u002F Tyr204）特异性的抗体对裂解物进行蛋白质印迹分析，并使用对p44 \u002F 42促分裂原活化蛋白激酶特异性的抗体进行适当的信号校正[1]。",{"key":257,"value":258},"细胞实验","人脐静脉内皮细胞（HUVEC）维持在M200 \u002F LSGS培养基中，细胞用于所有实验的第2代和第4代之间。对于增殖测定，将细胞以每孔3,000个接种在M200 \u002F 2％FBS中的0.1％明胶上4小时，然后在存在或不存在ATN-224的情况下用2ng \u002F mL FGF-2刺激直至48小时。使用Alamar Blue或MTT测定法测定HUVEC增殖。多发性骨髓瘤MM1S细胞在含有10％胎牛血清和2mM L-谷氨酰胺的RPMI 1640中生长。将HL-60早幼粒细胞白血病细胞和MOLT-4急性成髓细胞白血病细胞以400,000 \u002F mL接种在T-75烧瓶中，并孵育48至96小时用于增殖测定。使用钙黄绿素AM测定MMS1细胞增殖[1]。",{"key":260,"value":261},"动物实验","将小鼠[1]冷基质胶（500μL）与800ng \u002F mL FGF-2或300ng \u002F mL VEGF和肝素（50μg\u002F mL）混合。阴性对照栓塞不含促血管生成因子。将Matrigel混合物皮下注射到4至8周龄的雌性BALB \u002F c裸鼠中。在一些实验中，将具有或不具有Mn-TBAP（100μM）的ATN-224（94μM）或水分别直接添加至处理组和阴性对照组中的基质胶塞。或者，每周一至周五，每天用蒸馏水或ATN-224口服管饲小鼠。处死动物并在塞子注射后5天回收塞子。然后将塞子切碎并用组织匀浆器均化，并使用Drabkin溶液测定塞子中的血红蛋白水平。",{"key":263,"value":264},"参考文献","[1].\nJuarez JC, et al. 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