ZFIN Image: Haud et al., 2011, Fig. S4

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Figure Caption

Fig. S4 RNASET2 depletion in HEK 293 cells results in increased lysosome biogenesis without rRNA accumulation. (A) RNASET2 protein levels in HEK 293 cells transfected with control construct (control) or specific miRNA targeting RNASET2 detected by immunoblotting. (B) Cells transfected as in A for 48 h were stained with LysoTracker and fluorescence intensity measured in mCherry-positive cells using a flow cytometer (n = 3 independent experiments; error bars represent SEM). (C) EM examination of mCherry-positive cells transfected as in A for 48 h revealed more numerous lysosomes following RNASET2 knockdown. (Scale bar: 1 μm.) (D) Quantification of lysosome numbers from C. Mean number of lysosomes per EM grid quadrant measuring 500 μm² were enumerated (n = 3 independent experiments; error bars represent SEM; *P < 0.01, independent-samples t test). (E) Quantitative PCR was performed for 28S RNA: histogram depicts relative fold change in expression normalized to GAPDH ± SEM (n = 3).

Acknowledgments

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