Northern and In situ hybridization to confirm the probe is binding the junction site of chimeric HDAC11-S4 RNA 4.

A, Northern blotting for HDAC11-S4 RNA 4, partial HDAC11 RNA, and CPV S4 RNA. HDAC11-S4 RNA 4, partial HDAC11 RNA, and CPV S4 RNA prepared by in vitro transcription were separated on 1% agarose-formaldehyde gels and transferred to Hybond-N+ nylon membranes. Northern blotting was conducted with biotin-labeled DNA targeting the junction site of chimeric HDAC11-S4 RNA 4. B, In situ hybridization of Ctenopharyngodon idellus kidney (CIK) cells respectively transfected with HDAC11-S4 RNA 4, partial HDAC11 RNA, and CPV S4 RNA. A total of 1×10⁴ cultured CIK cells seeded in 24-well plates, and followed by transfection with partial HDAC RNA (2μg), BmCPV S4 RNA (2μg) or HDAC11-S4 RNA 4 (2μg). The cells were collected at 48 h post transfection and digested with proteinase K and fixed in 4% formaldehyde at 4°C for 1 h. The cells were hybridized with the biotin-labeled DNA targeting the junction site of chimeric HDAC11-S4 RNA 4. CY3-labeled streptavidin was used to visualize hybridization signals. Nuclei were stained with DAPI.

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