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. 2009 Jun 1;20(11):2744–2754. doi: 10.1091/mbc.E08-11-1092

Figure 2.

Figure 2.

Inhibition of lysosomal degradation in Hsp47−/− cells results in accumulation of collagen in lysosomes. Hsp47+/+ (A–C) and Hsp47−/− (KO-13) cells (D–F) were cultured in the presence (+) or absence (−) of E64d/pepstatin A. Localization of type I collagen was analyzed by double staining with antibodies against localization marker proteins. PDI (A and D), GM130 (B and E) and LAMP2 (C and F) were used as ER, Golgi, and lysosome markers, respectively. Bar, 10 μm. (G) Preembedding immunoelectron microscopic observation of type I collagen in Hsp47−/− cells under lysosome inhibition conditions. Cells were treated with E64d/pepstatin A. An arrow indicates amphisomes-like structures. Bar, 500 nm. (H) Double-labeling immunoelectron microscopy of type I collagen and LAMP-2 in Hsp47−/− cells. Type I collagen and LAMP-2 are labeled with gold particles of 10 nm (arrows) and 5 nm (arrowheads), respectively. (I) A high-magnification view of the region boxed in H. Bar, 100 nm.