Journal: Cell Death and Differentiation

Article Title: The MRN complex is transcriptionally regulated by MYCN during neural cell proliferation to control replication stress

doi: 10.1038/cdd.2015.81

Figure Lengend Snippet: Mycn promotes MRN complex expression in Hh-induced mouse cerebellar GCPs. ( a and b ) WB and real-time Q-PCR analysis of the expression of the indicated proteins/transcripts in mouse cerebella at the indicated post-natal (P) days; Zic1 and gamma-aminobutyric acid receptor subunit alpha-6 (Gabra6) were used as differentiation markers and the ponceau staining was used as the loading control. For Q-PCR, the levels of mRNAs were normalized to Hprt expression and reported as fold variation compared with P5 levels. ( c ) β -tubulin immunofluorescent staining and WB analysis of Mycn and MRN complex proteins in GPC cultures (DIV: days in vitro ). ( d ) Q-PCR assessment of the expression of the indicated transcripts normalized as above and expressed as fold variation compared with the levels at DIV 1. ( e – g ) WB ( e and g ) and real-time Q-PCR analysis ( f ) of the expression of the indicated proteins/transcripts in GPCs treated with DMSO or SAG for 48 h without ( e – g ) or with ( g ) lentiviral-transduced anti-Mycn shRNAi

Article Snippet: RNA interference experiments were all performed using validated small interfering RNA sequences (Sigma Aldrich) transfected by Dharmafect 2 Reagent (Thermo Fisher Scientific, Waltham, MA, USA) or by means shRNA lentiviral transduction particles (Santa Cruz Biothecnology, Dallas, TX, USA and Sigma Aldrich).

Techniques: Expressing, Staining, Control, In Vitro