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. 2018 Oct 8;15(3):438–452. doi: 10.1080/15548627.2018.1525476

Figure 4.

Figure 4.

LC3BY113C mutation reduces physical interaction with ATG7, but not ATG3. (a) The relative interaction of GLuc (Fr1)-LC3B with ATG3-GLuc (Fr2) and ATG7-GLuc (Fr2) measured by PCA. The results calculated are the mean of 3 independent experiments and normalized to the GLuc (Fr1)-LC3B WT variant. Statistical significance determined by the two-tailed unpaired Student t test, **P < 0.01. (b) Western blot of the PCA samples, blotted with anti-Gaussia antibodies. VCL was used as a loading control. (c) Co-immunoprecipitation of HA tagged LC3B variants with endogenous ATG3 and ATG7. VCL was used as loading control for total cell extract. (d) The relative coIPed levels of endogenous ATG3 and ATG7 with LC3B variants (LC3B WT and LC3BY113C). IP levels were quantified using densitometry and coIPed endogenous ATG7 and ATG3 levels were normalized to IPed HA-LC3B variants. The results calculated are the mean of 3 independent experiments and normalized to the HA-LC3B WT variant. Statistical significance determined by the two-tailed unpaired Student’s t-test, using Holm-Sidak correction, *P < 0.05; **P < 0.01.