FIGURE

Fig. 6

ID

ZDB-FIG-210217-61

Publication

Turan et al., 2020 - The endosomal trafficking regulator LITAF controls the cardiac Nav1.5 channel via the ubiquitin ligase NEDD4-2

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Fig. 6

Figure 6. LITAF causes ubiquitination and subsequent proteasomal degradation of NEDD4-2 in HEK cells. A, IP of lysates from HEK cells transfected with plasmids for FLAG-tagged NEDD4-2, HA-tagged ubiquitin, FLAG-tagged LITAF, or control plasmid for 48 h was performed with anti-HA antiserum. A representative immunoblot shows levels of ubiquitinated NEDD4-2–FLAG and tubulin and input levels of NEDD4-2–FLAG, LITAF-FLAG, or tubulin. Relative NEDD4-2–FLAG ubiquitination levels as calculated by the ratio of ubiquitinated NEDD4-2 normalized to ubiquitinated tubulin and total NEDD4-2 normalized to total ubiquitin levels (n = 5; means ± S.E.). In the scatter plot, averaged values for the five independent experiments together with the means and S.E. values are shown. Bottom panel, Student's t test, p < 0.01. B, LITAF-mediated degradation of NEDD4-2 through proteasomes. HEK cells were transfected with plasmids for NEDD4-2–FLAG, LITAF-HA or control plasmid for 24 h and then treated with vehicle (▿), 5 μm MG132, or 10 μm chloroquine for 24 h. Top panel, representative Western blots show total expression of NEDD4-2–FLAG, LITAF-HA, and tubulin of treated cells. Bottom panel, respective relative expression levels (means ± S.E.) of total NEDD4-2 normalized to tubulin levels (n = 4, n = 2, p < 0.05).

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

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