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Different parts of Ficus religiosa tree have traditional usage against many ailments and diseases. In the present study, 85% ethanol extract of F. religiosa leaves were partially purified by PLC Silica gel 60 F254, 2 mm, 20X20 cm preparative thin layer chromatography using toluene and ethyl acetate (8:2, v/v) as solvent. Fraction 7 (FR7) showed highest brine shrimp larvicidal activity as regard to the leaf extract of F. religiosa. In vitro anticancer potential of same fraction against Jurkat cell line was investigated by means of flow cytometry using annexin V/7AAD apoptosis kit. F. religiosa leaf extract was found to markedly inhibit the proliferation of Jurkat cells in a dose-dependent manner.