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. 2016 Sep 8;7(46):74612–74629. doi: 10.18632/oncotarget.11916

Figure 9. GFP-LC3 puncta formation assay confirms Fyn's regulation of AICAR dependent autophagy.

Figure 9

A. HEK293T cells were transfected with both siRNA for Fyn and GFP-human LC3 construct. The cells were treated with 10 ng/ml TNFα for 36 h with or without 2 mM AICAR for 10 min. Cells were fixed with 4% paraformaldehyde and mounted with DAKOCytomation Fluorescent Mounting Medium (S3023). These are representative images of high power field from experiments independently performed 3 times. B. Number of puncta in each single cell. Data are representative of n = 5 experiments. Statistical examination between control and AICAR-stimulated cells in each sets were performed ***P < 0.005. C. HEK293T cells were transfected with both siRNA for Fyn and GFP-human LC3 construct. Cells were treated with 10 ng/ml TNFα for 36 h with or without 100 nM Bafilomycin for 4 h and 2 mM AICAR for 10 min. Cells were fixed with 4% paraformaldehyde and mounted with DAKOCytomation Fluorescent Mounting Medium (S3023). These are representative images of high power field from experiments independently performed 3 times. D. Number of puncta in each single cell. Data are representative of n = 5 experiments. Statistical examination between control and AICAR stimulated cells in each sets were performed ***P < 0.005.