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Fig. 4 | Journal of Neuroinflammation

Fig. 4

From: Neuroinflammation driven by human immunodeficiency virus-1 (HIV-1) directs the expression of long noncoding RNA RP11-677M14.2 resulting in dysregulation of neurogranin in vivo and in vitro

Fig. 4

Overexpression of RP11-677M14.2 suppresses Nrgn expression. (A) HEK293T cells were transiently transfected with pCDNA3.1 RP11-677M14.2 (pCDNA 3.1 Nrgn-AS) and after 48 h the levels of Nrgn mRNA were assessed by RT-qPCR in comparison with cells transfected with empty vector. (B) HEK293T cells were transfected with different doses of pCDNA3.1 Nrgn-AS as indicated, and levels of Nrgn mRNA were assessed after 48 h by RT-qPCR and compared with empty vector. Levels of (C) Nrgn mRNA and (D) protein were assessed by RT-qPCR and Nrgn ELISA respectively upon stably transfection of SH-SY5Y with pCDNA3.1 Nrgn-AS compared to empty plasmid (N = 3). (E-F) Representative images of single molecule RNA HCR FISH with probe for RP11-677M14.2 transcript (AF-488, green puncta) in undifferentiated stably transduced SH-SY5Y cells compared to SH-SY5Y transduced with empty vector. DAPI stain for cell nuclei is shown in blue. Scale bars show 10 μm, N = 3. Effect of RP11-677M14.2 knockdown on Nrgn expression in (G) HEK293T and (H) SH-SY5Y cells assessed by RT-qPCR 72 h after transduction (N = 4)

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Journal of Neuroinflammation

ISSN: 1742-2094