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. Author manuscript; available in PMC: 2021 Sep 27.
Published in final edited form as: N Engl J Med. 2021 Jun 24;384(25):2406–2417. doi: 10.1056/NEJMoa1915722

Figure 5. Complementation Studies with Fibroblasts and a Summary of Findings.

Figure 5

Panel A shows the results of transient introduction of plasmids encoding wild-type ATG7 into immortalized fibroblasts from Patient 1 for 24 hours. Cells were then treated with or without CQ (60 μmol per liter) and AZD8055 (1 μmol per liter) for 2 hours before immunoblot detection of ATG7, LC3, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, loading control). The Western blots shown are representative of triplicate experiments. Panel B shows the results of transient introduction of plasmids encoding wild-type, catalytic-null (C572A), and missense ATG7 variants into Atg7-knockout mouse embryonic fibroblasts for 24 hours. Cells were then treated with CQ (60 μmol per liter) and AZD8055 (1 μmol per liter) for 2 hours. Immunoblotting results are representative of three independent experiments with antibodies against ATG7 and LC3. GAPDH was used as a loading control. The graph in Panel B shows the results of a densitometric analysis of the ratio of LC3 to GAPDH, normalized to wild-type values. The bars represent means, and T bars indicate standard deviations. Panel C shows a summary of our findings.