|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
|
Fig. 2. FHL3 overexpression retards myogenin induction following 24 hours differentiation. (A) Lysates from differentiating C2C12 myoblasts stably expressing Vector or HA-FHL3 were immunoblotted for MyoD, myogenin or total actin using specific antibodies. Immunoblots are representative of three independent experiments. (B) The mean relative level of myogenin protein in C2C12 myoblasts stably expressing Vector or HA-FHL3 after 24 hours of differentiation was quantified by densitometry and presented as the combined data from Vector or HA-FHL3 (1) and (2) cell lines (n=3, **P<0.01). The myogenin protein levels in vector cell lines standardised to actin were arbitrarily set at 1 and myogenin levels in HA-FHL3 lines expressed relative to vector cell lines. Error bars represent ± s.e.m. (C) C2C12 myoblasts were differentiated for 24 hours, stained with anti-myogenin antibodies (blue), and propidium iodide (red) and imaged by confocal microscopy. Bar, 20 µm. Yellow arrowheads indicate myogenin-positive nucleus. White arrowheads indicate myogenin-negative nucleus. Shown are representative images for Vector (2) and HA-FHL3 (2) cell lines. Vector (1) and HA-FHL3 (1) cell lines showed equivalent results (data not shown). Results shown are representative of three independent experiments. (D) The mean percentage of nuclei expressing myogenin was quantified following 24 hours of differentiation and is presented as the combined data from Vector and HA-FHL3 (1) and (2) cell lines. A minimum of 116 nuclei were scored for each stable cell line per time point for each of three independent experiments (**P<0.01). Error bars represent ± s.e.m.
|
