Figure 3

Collagen type VI (ColVI) expression study in cultured skin fibroblasts. (A) Representative images obtained using the protocol from [67] in which ColVI (red) is labeled with monoclonal antibody MAB1944 (Chemicon (now Millipore), Billerica, MA, USA) and perlecan (green) with monoclonal antibody MAB1948 (Chemicon). Note that ColVI expression appeared clearly altered in a patient with an early severe (ES) form and less so in patients with intermediate (Int) or Bethlem myopathy (BM) forms, compared with control fibroblasts (Cont). (B) Using the protocol of Hicks et al. [64], which detects ColVI (red) with polyclonal antibody Ab6588 (Abcam, Cambridge, UK) and fibronectin (green) with monoclonal antibody F15 (Sigma Chemical Co., St Louis, MO, USA), the sensitivity of the method is increased, and defective ColVI secretion could be detected in all patients' samples. Insets indicate nuclei, labeled using DAPI. Bars are 50 μm. (Images courtesy of Corine Gartioux and Valérie Allamand).