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Figure 4

ID
ZDB-IMAGE-200130-18
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Figures for Chen et al., 2019
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Figure 4

LPA3 is highly internalized and sorted to the lysosome degradation pathway in Progerin cells. (a) Western blot results show that treating Progerin HEK293 cells with 5 μM of MG132 for 6 hr showed no LPA3 protein restoration. DMSO was used as solvent control (Ctrl). (b) Western blot results show that Progerin HEK293 cells were treated with 5 mM of NH4Cl or 20 nM of Bafilomycin A1 (BA‐1) for 6 hr to block LPA3 degradation through the lysosome pathway. ddH2O was used as solvent control for NH4Cl (Ctrl in upper panel). DMSO was used as solvent control for BA‐1 (Ctrl in lower panel). (c) Western blot results show that HGPS AG11 fibroblasts were treated with 5 mM of NH4Cl and 5 μM of MG132 for 6 hr. NH4Cl treatment rescued LPA3, but not LPA1 or LPA2, in HGPS AG11 fibroblasts. ddH2O and DMSO were both loaded to control sample as solvent control (Ctrl). (d) Counter‐immunofluorescent staining by LPA3 and LAMP‐1 shows that LPA3 localized into the lysosomes in Progerin HEK293 cells. (e) Western blot results show that NH4Cl treatment rescued LPA3 in HGPS AG03 fibroblasts. ddH2O and DMSO were both loaded to control as solvent control (Ctrl). (f) Scheme representing the internalization assay of LPA receptors. (g) Representative images of time‐dependent LPA3 internalization. Internalization of LPA3 was higher in Progerin HEK293 cells. (h) Integrated density of intracellular fluorescence was quantified to indicate internalized LPA3. ANOVA and Student's t test; **p < .01, ***p < .001

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