Fig. 5

Increased lysosomal hydrolytic capacity induced by chronic or acute AEP-deficiency is controlled by the Jak2-STAT3 signalling pathway. a STAT3 activation in WT but not AEP^−/− MEFs treated for 16 h with increasing concentrations of MVO26630. Data are the average ± SD of n = 3 biologically independent samples. Statistical significance was calculated using a Dunnett’s multiple comparison test. b P-STAT3 localisation in WT and AEP^−/− MEFs treated or not with 50 μM MVO26630 for 16 h (endogenous P-STAT3) and HKC-8 cells transfected with STAT3-YFP and treated or not with 50 μM MVO26630 for 16 h. Data are the average ± SEM of n = 96 cells for control samples and n = 86 for MVO-treated samples. Statistical significance was calculated using a two-sided unpaired t-test. (Scale bar = 20μm). c STAT3 activation in HKC-8 cells treated with increasing levels of MVO26630 overnight. Data are the average ± SD of n = 3 biologically independent samples. Statistical significance was calculated using a two-sided unpaired t-test. d CtsD, CtsB and P-STAT3 levels in WT MEFs compared to WT MEFs treated overnight with 50 μM MVO26630 with or without 100 μM of the STAT3 inhibitor S3I-201. Data are the average ± SD of n = 3 biologically independent samples. Statistical significance was calculated using a two-sided unpaired t-test. e Levels of STAT3, CtsB and CtsD in WT HKC-8 cells transfected with a non-targeting siRNA (NT) or two different STAT3-targeting siRNAs (#8 and #10) and treated or not with 50 μM MVO26630 for 16 h. Data are the average ± SD of n ≥ 3. Statistical significance was calculated using a two-sided unpaired t-test. f P-STAT3 western blot in WT HKC-8 treated with 50 μM MVO26630 in the presence or absence of the Jak2 inhibitor SD-1029 (10 μM). Data are the average ± SD of n = 3 biologically independent samples. Statistical significance was calculated using a Dunnett’s multiple comparison test. g Levels of P-Jak2 in WT HKC-8 cells treated or untreated with 50 μM MVO26630 for 16 h. Data are the average ± SD of n = 3 biologically independent samples. Statistical significance was calculated using a two-sided unpaired t-test. h Levels of Jak2, P-STAT3, CtsB and CtsD in WT HKC-8 cells transfected with a non-targeting siRNA (NT) or two different Jak2-targeting siRNAs (#6 and #7) and treated or not with 50 μM MVO26630 for 16 h. Data are the average ± SD of n ≥ 3 biologically independent samples. Statistical significance was calculated using a two-sided unpaired t-test