Figure 3.

ANG is conventionally secreted under ER stress. (A) Schematic representation of the ANG protein sequence. The functional domains are annotated in dark gray. (B) Box-and-whisker plots representing ANG, Binding immunoglobulin protein (BiP), CCAAT-enhancer-binding protein homologous protein (CHOP) and IL-6 transcripts relative expression analyzed by RT-qPCR in HREC incubated for 16 hours with 5 μg/ml BFA or vehicle. Data are from three independent experiments. The dotted line represents 1, the expression level of transcripts in the control condition when measured by the 2^{(–∆∆CT)} method. (C,D) Box-and-whisker plots representing (C) IL-6 and(D) ANG concentrations measured by ELISA in the extracellular medium of HREC incubated with 2.5 μg/ml tunicamycin or vehicle and coincubated with 5 μg/ml BFA for 24 hours. Data are from three independent experiments. Mann–Whitney U test: *P<0.05. (E) Box-and-whisker plots representing ANG transcripts relative expression analyzed by RT-qPCR in HREC incubated for 16 hours with 5 μg/ml BFA, 2.5 μg/ml tunicamycin, both, or vehicle. Data are from three independent experiments. The dotted line represents 1, the expression level of transcripts in the control condition when measured by the 2^{(–∆∆CT)} method. (F) Immunofluorescence analysis by confocal microscopy of ANG expression in HREC incubated for 8 hours with 2.5 μg/ml tunicamycin, 0.25 μM thapsigargin, 5 μg/ml BFA, or vehicle. The bar represents 10 μm.