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. 2019 Dec 12;94(1):e01311-19. doi: 10.1128/JVI.01311-19

FIG 6.

FIG 6

The CC2 domain of HRS suffices to stabilize APH-2, and an EXXXLL motif in APH-2 targets it for lysosomal degradation. (A and B) Cellular lysates of HEK293T expressing FLAG-APH-2 plasmid and Myc-HRS (A) or HA-HRS (B) deletion mutants were analyzed by Western blotting using anti-FLAG, anti-Myc, anti-HA, and anti-α-tubulin antibodies. Results are shown as fold changes in comparison with the condition in which APH-2 was expressed alone, which was arbitrarily set as 1. Error bars represent the standard error of the mean. *, P ≤ 0.05; **, P ≤ 0.01, by two-tailed Student's t test. Data represent three independent experiments. (C) The effect of the EXXXLL motif on APH-2 stabilization was assessed. HeLa cells were transfected with the indicated FLAG-APH-2 mutants and an empty control plasmid or HA-HRS expression plasmid. Cellular lysates were analyzed by Western blotting using anti-FLAG and anti-α-tubulin antibodies. The value for APH-2 expressed alone was arbitrarily set as 1, and error bars represent the standard error of the mean. Data represent two independent experiments.