Fig. 8: RBPJ is a potential transcription factor of ORC6 in glioma cells.

Multiple databases predicted the common potential transcription factors of ORC6 (A). P1 glioma cells were transfected with the specified siRNAs (at 200 nM) targeting the described transcription factors or a scramble non-sense siRNA (siC, at 200 nM) for 48 h, the expression of ORC6 mRNA was assessed (B). P1 glioma cells were genetically engineered to stably express the lentiviral RBPJ shRNA (shRBPJ-Sq1 or shRBPJ-Sq2) (C and D), scramble control shRNA (“shC”) (C, D), the lentiviral RBPJ-expressing construct (oeRBPJ) (E, F), or an empty vector (“Vec”) (E, F); The expression of the listed mRNAs and proteins was assessed (C–F). Chromatin Immunoprecipitation (ChIP) assay results showed the relative levels of RBPJ-bound ORC6 promoter region in specified glioma tissues (“T”) and matched adjacent normal brain tissues (“N”) (G) as well as in listed glioma cells and human astrocytes (H). RBPJ protein expression in primary human astrocytes (“Astrocytes1/2”), immortalized (A172) or primary (“P1,” “P2,” “P3”) glioma cells was presented (I). Values were mean ± standard deviation (SD). *P < 0.05 versus “siC” (B). *P < 0.05 versus “shC”/ “Vec” cells (C–F). *P < 0.05 versus “N” tissues or “Astryocytes1” (G, H). “n. s.” indicates non-statistical difference (P > 0.05). The experiments were conducted five times (biological repeats), yielding consistent results.