Fig. 2: JNK inhibition impacts on lysosomal homeostasis. | Cell Death Discovery

Fig. 2: JNK inhibition impacts on lysosomal homeostasis.

From: Inhibition of JNK increases the sensitivity of hepatocellular carcinoma cells to lysosomotropic drugs via LAMP2A destabilization

Fig. 2

A Cytofluorometric analysis of intracellular acidification using acridine orange in Hep3B pre-treated for 6 h with 10 μM SP600125 and then treated with 300 μM GPN for an additional 2 h. Data are expressed as mean ± SEM of n = 3 independent experiments. ***p < 0.001 vs. CTRL; §p < 0.05 vs. DMSO. B Fluorescence microscopy analysis of galectin-3 puncta in Hep3B pre-treated for 6 h with 10 μM SP600125 and then treated with 300 μM GPN for an additional 2 h. Data are expressed as mean ± SEM of n = 3 independent experiments. **p < 0.01 vs. CTRL; §§p < 0.01 vs. DMSO. C Western blot analysis of the release of cathepsin-B in the cytosol in Hep3B pre-treated for 6 h with 10 μM SP600125 and then treated with GPN for an additional 2 h. LAMP1 and GAPDH were used as controls of fraction purity. D and E Western blot analysis of PARP1 in Hep3B treated for 24 h with the indicated concentration of the lysosomotropic agents GPN (D) and chloroquine (E) in the presence or absence of 10 μM SP600125. F Western blot analysis of PARP1 in Hep3B overexpressing LAMP2A and treated for 24 h with the indicated concentration of the lysosomotropic agents GPN in the presence or absence of 10 μM SP600125. Where applicable, tubulin was used as a loading control. Western blots are representative of n = 3 independent experiments showing similar results.

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