Figure 6.

Fibroblasts Have Abnormal Golgi Morphology and Impaired Autophagic Flux

(A and B) Fibroblasts from affected individuals and a control were cultured at 37°C or 40°C and show (A) an expanded Golgi network with proband 1 also showing fragmentation of the Golgi at 40°C, compared to control by GM130, and (B) possible expansion of the ER-Golgi intermediate compartment (ERGIC) by ERGIC53.

(C) Fibroblasts cultured with and without bafilomycin A1 (BafA1) show increased LC3 by immunofluorescent imaging that was not further increased by bafilomycin A1 treatment compared to control.

(D) Whole-cell lysates from each proband show an increase in LC3-II/Actin ratio compared to control. The LC3-II/Actin ratio in control cells was increased with bafilomycin A1 treatment, but remained unchanged in probands 1 and 3.

Ctrl, control; Pr, proband; bars represent mean ± standard deviation for three independent experiments. Two-tailed paired t test.