Fig. 2: Nedd4-1(NE) localization is regulated by essential amino acids. | Nature Communications

Fig. 2: Nedd4-1(NE) localization is regulated by essential amino acids.

From: The primate-specific Nedd4-1(NE) localizes to late endosomes in response to amino acids to suppress autophagy

Fig. 2

HeLa cells were transfected with 3xFlag-Nedd4-1(NE) and incubated for 1 hr in (A) DMEM (supplemented with 10% serum), (B) HBSS, or (C) HBSS supplemented with an essential amino acid (EAA) cocktail plus glutamine (Gln; 4 mM). Cells were fixed, permeabilized, and immunostained with anti-Flag antibodies (green). Nuclei were stained with DAPI (blue). D–J Nedd4-1(NE) localization was scored blindly as either “diffuse - cytoplasm/membrane”, “punctate”, or “both”, as indicated by the examples in (H–J). Scale bars are (A–C) 10 μm, or (H–J) 15 μm. Data are means ± SEM of 3 independent experiments. Statistical significance was determined using a chi-squared test (N.S.: not significant; **: p < 0.01; ****: p < 0.0001). In (E), statistical tests are comparing each condition to HBSS. Cell counts for each condition are indicated in Table S2.

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