Figure 7

Jam-a expression and lysosomal Jam-a accumulation upon cysteine cathepsin inhibition. (A) Western Blot of Jam-a in whole cell lysates of untreated −/+E64d and four days TGFβ-1 −/+E64d treated iPL32 cells showed full length (FL) Jam-a and a 25 kDa Jam-a fragment (F1). (B) Jam-a transcription was analyzed by qRT-PCR in untreated and four days TGFβ-1 −/+E64d treated iPL32 cells. Starting quantity values were normalized to the untreated group and are shown as the mean ± SEM (n = 3). (C) Representative confocal images of FITC-Phalloidin (green) and Jam-a (red) immunofluorescence staining of untreated and four days TGFβ-1 −/+E64d treated iPL32 cells are shown. Images represent one confocal section at a medial position in the cells. (D) Representative images of optical sections of Jam-a/Lamp-1 immunofluorescence double-staining of four days TGFβ-1 −/+E64d treated iPL32 cells: Jam-a (red), Lamp-1 (green), and nuclei (blue) are shown. Dashed lines mark areas shown in higher magnification in the second, third, and fourth panels. Arrows denote Jam-a co-localization with Lamp-1 positive lysosomes. Arrowheads indicate Jam-a staining adjacent to Lamp-1 positive lysosomes. Scale bars = 10 μm.