Collagen IV accumulates intracellularly and exhibits decreased stability and molecular mass in Colgalt1 mutant MEFs. Immunoblot detection of the α2 chain of type IV collagen in lysates (A) and conditioned media (B) from wild-type and Colgalt1fosse/fosse MEF cultures. Ponceau S staining is shown as a loading control. Arrows indicate the band corresponding to full-length collagen IV. Graphs show densitometric quantitation of COL4A2 signal intensities normalized to Ponceau S. (C) Immunoblot detection of monomer and dimers of the NC1 domain of COL4A2 in collagenase digests of the ECM isolated from wild-type and fosse MEFs. The graph shows densitometric quantitation of the total (monomers and dimers) signal intensity from five different samples in each MEF group. Results are shown as mean (s.d.) and P-values ≤0.05 and ≤0.01 are indicated by one (*) or two (**) asterisks, respectively. (D) Immunofluorescent detection of type IV collagen in muscle tissue from wild-type and fosse mice. Magnification: 40×.
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