Fig. 1 | Chromosoma

Fig. 1

From: Ectopic expression of pericentric HSATII RNA results in nuclear RNA accumulation, MeCP2 recruitment, and cell division defects

Fig. 1

Transient transfection of satellite expression results in nuclear satellite RNA accumulation. (a) Transfection scheme for transient and stable integration expression. A plasmid harboring HSATII, α-sat (αsat), or no insert (empty vector) is introduced to cultured HeLa or Tig-1 primary fibroblast cells via lipid-mediated transfection and cells are then fixed on coverslips or harvested for RNA isolation. Stable cell lines are further selected with neomycin (G418) for 2 weeks prior to fixation or harvesting. Twenty-four hours after transfection, nuclei are scored for expression of (b) HSATII and (c) α-sat RNA signal by RNA FISH. Percent of cells (out of 500 nuclei) with (d) HSATII RNA nuclear expression and (e) α-sat nuclear expression. (f) Nuclear RNA signal detected by RNA FISH is dependent on the sequence harbored within the transfected construct. Asterisks denote significant differences from empty vector transfected cells by Chi-square test, p<.0001. (g) qRT-PCR of HSATII RNA 24 h after transfection. Expression values shown relative to housekeeping gene expression (ß-actin) for each transfection condition. Scale bar, 5μm

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