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. 2013 Aug 9;41(20):9424–9437. doi: 10.1093/nar/gkt717

Figure 6.

Figure 6.

Gene knockdown of EhMyb1 reveals that AS sRNAs are generated to nascent transcript. (A) N-terminal Myc-tagged EhMyb has nuclear localization. (B) E. histolytica HM-1:IMSS parasites stably transfected with the plasmid containing the EHI_197520 trigger fused to the EhMyb1 gene (cell line: 19-trigger-Myb). RT-PCR for EhMyb1 in the 19-trigger-Myb cell line demonstrates the absence of EhMyb mRNA in the 19-trigger-Myb cell line. Control (EHI_199600) indicates equivalent mRNA in both lanes. (C) Northern blot of the 19-trigger-Myb cell line probed for EhMyb1 AS sRNAs reveals an abundant population of AS sRNA to the EhMyb1 gene. AS sRNAs that map to the intron of EhMyb1 were detected. Control (EHI_118130) indicates equivalent sRNA in both lanes (75 µg of enriched sRNA). (D) Viability assay of stress-treated parasites (1 h, 1 mM H2O2) revealed that EhMyb1 knockdown parasites had significantly decreased viability in response to stress.