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Silkworm Piwi proteins repressed transcription in an HP1-dependent manner.

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posted on 17.03.2014, 03:13 by Tsuneyuki Tatsuke, Li Zhu, Zhiqing Li, Hitoshi Mitsunobu, Kaito Yoshimura, Hiroaki Mon, Jae Man Lee, Takahiro Kusakabe

(A–F) Transcriptional repression of luciferase 2P (luc2P) reporter gene under the control of the hsp promoter by silkworm Piwi and HP1 proteins in BmN4-SID1 cells with dsRNA against Ago3, Siwi, HP1a or HP1b. The hsp promoter-based luc2P reporter contains five synthetic GAL4-UAS sites and was used in all transfections. BmN4-SID1 cells were transfected with a Luc2P reporter under the control of the hsp promoter, upstream of which five UAS were located for the hsp promoter. Cells were co-transfected with an expression vector for GAL4 DNA-binding domain (DBD)-fused Ago3, Siwi, HP1a or HP1b proteins at 72 h after dsRNA introduction. Luciferase activities were measured at 72 h post-transfection. Luciferase activities are represented as relative values based on DBD (empty plasmid introduced cells) as a standard. Error bars = SD. The SDs and P-values (determined by the t-test, *P<0.1, **P<0.05, ***P<0.01, which was checked by a comparison with the luciferase activities in the DBD-introduced controls with DBD-HP1a, DBD-HP1b, DBD-Ago3 or DBD-Siwi-introduced cells) are based on n = 3.