Figure 3
PR-LncRNA1 and 10 silencing leads to increased proliferation and stemness. U87-MG cells were transfected with specific ASOs for the PR-LncRNAs indicated. (A,B) Transfected cells were examined for PR-LncRNA1 and PR-LncRNA10 expression by quantitative reverse transcription polymerase chain reaction (n = 4). (C) Representative immunofluorescence of P-H3 in U87MG cells under the conditions indicated. (D) Quantification of the number of P-H3 positive cells under the conditions indicated (n = 4). (E) Quantification of mRNA levels of p21cip, Bax and SerpinB5 in cells transfected with ASOs for PR-LncRNA1 and 10 and compared to cells with a control ASO (F) Expression of PR-LncRNA 1,5 and 10 in indicated conventional cell lines (U87-MG, U251, U373 and A172) and glioma stem cells (GNS166, GNS179 and GB1) (G) Quantification of primary oncospheres formed in ASO-transfected cells after 10 days in culture (n = 3). (H) Quantification of number of secondary oncospheres generated from disaggregating primary oncospheres in ASO-transfected and control cells. Numbers were assessed after 10 days in culture (n = 3). Asterisks (*,**and ***) indicate statistical significance (p < 0.05, p < 0.01, and p < 0.001, respectively).
