Journal: Cell Death and Differentiation

Article Title: Haploinsufficiency of GCP4 induces autophagy and leads to photoreceptor degeneration due to defective spindle assembly in retina

doi: 10.1038/s41418-019-0371-0

Figure Lengend Snippet: Autophagy flux associated with GCP4. a Detection of autophagy flux using fluorescent confocal microscopy. Stable miR- Gcp4 -2# and miR- LacZ COS-7 cells were transfected with a tandem expression vector mCherry-GFP-LC3 and cultured in normal and EBSS medium for 2 h. In the BAF + EBSS group, the cells were treated with BAF for 4 h to suppress the fusion between autophagosome and lysosome. Yellow or green puncta indicated autophagosomes, while red puncta include autophagosomes and autolysosomes, because GFP protein is sensitive and attenuated in an acidic environment of autolysosome. Scale bar: 10 μm. b The tandem structure of mCherry-GFP-LC3 and statistical analysis of LC3 puncta per cell ( n = 3 experiments, with 40 cells/experiment). Data were represented as means ± SD. Two-way ANOVA followed by Bonferroni posttest was used for statistical analysis. * p < 0.05; ** p < 0.01

Article Snippet: For starvation treatment, the cells were cultured in EBSS medium (SH30029.02, HyClone).

Techniques: Confocal Microscopy, Transfection, Expressing, Plasmid Preparation, Cell Culture