Journal: PLoS ONE

Article Title: Clathrin adaptor GGA1 modulates myogenesis of C2C12 myoblasts

doi: 10.1371/journal.pone.0207533

Figure Lengend Snippet: ( A ) Myogenic differentiation of C2C12 cultured myoblasts. C2C12 cells cultured in growth medium (day 0) and shifted to differentiation medium for four days (day 4) were stained with anti-myosin heavy chain 3 antibodies. ( B ) C2C12 cells cultured in the growth medium (day 0, a-o) and differentiation medium for four days (day 4, a′-o′) were subjected to immunofluorescent microscopy using anti-golgi associated, gamma adaptin ear containing, ARF binding protein 1 (GGA1) (a, c, d, f, g, i, m, o, a′, c′, d′, f′, g′, i′, m′, and o′), anti-golgin subfamily A member 2 (GM130) (b, k, b′, and k′), anti-adaptor related protein complex 1 subunit gamma 1 (AP1G1) (AP1: e, n, e′, and n′), anti-Ras-related protein rab-4A (RAB4) (h and h′), anti-clathrin heavy chain (CHC) (j and j′), and anti-lysosomal associated membrane protein 1 (LAMP1 (n and n′) antibodies. The cells from day 0 were stained with anti-GGA2 (p and r) and anti-AP1G1(q and r). All the images from anti-GGA1 or GGA2 antibodies are shown in green and those from other antibodies are shown in red. Merged images are indicated. Scale bar: 10 mm.

Article Snippet: The primary antibodies used were as follows: anti-GGA1 or 2 (this study), anti-Golgin subfamily A member 2 (GM130), anti-Ras-related protein rab-4A (RAB4), anti-clathrin heavy chain (CHC), and anti-adaptor related protein complex 1 subunit gamma 1 (AP1G1) antibodies were from BD Biosciences (San Jose, CA, USA), whereas anti-lysosomal associated membrane protein 1 (LAMP1; 1D4B) rat monoclonal antibody, anti-MYH3, and anti-beta actin antibodies were purchased from Santa Cruz (Dallas, TX, USA).

Techniques: Cell Culture, Staining, Microscopy, Binding Assay