Fig. 3

miR-449a binding to Jag1 inhibits Notch target genes and NICD levels. a Differentiated C2C12 cells were transfected with the miR-449a mimic (10 nM) in the absence or presence of its inhibitor (10 nM). After 48 h, cells were lysed and relative transcript levels of Notch target genes, Hes1 (a) and Hey1 (b) were quantified by qRT-PCR and normalised to 18S rRNA. Control cells were transfected with the scramble (Scr). c Differentiated C2C12 cells were grown on six-well plates and transfected with miR-449a (10 nM) alone or in the presence of its inhibitor (10 nM). After 48 h, cells were fixed in 4% formaldehyde and immunofluorescence was performed with anti-NICD antibody as described in ‘Materials and Methods’ (d) Human primary skeletal muscle cells were grown on chamber slides and transfected with either scramble (Scr) or miR-449a alone or with its inhibitor (10 nM). As in (c), immunofluorescence was performed and cells were visualised in fluorescence microscope. Incubations were repeated at least three times and values are presented as mean ± SEM. ***p < 0.001, *p < 0.05