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. 2017 Nov 8;8:1507. doi: 10.3389/fimmu.2017.01507

Figure 3.

Figure 3

Leptin decreases endoplasmic reticulum (ER) stress-induced autophagy flux by reducing LC3II turnover and SQSTM1 degradation in mice adipocytes. (A) Representative fluorescent photomicrographs showing the GFP-LC3 puncta formation in adipocytes transfected with GFP-LC3 plasmid and stained with Lyso-tracker Red. Cells were treated with tunicamycin (TM) or leptin. The nuclei were stained with DAPI shown in blue (n = 3). (B) Representative electron micrographs (30,000×) of adipocytes. Yellow arrowhead: endoplasmic reticulum, red arrowhead: mitochondria, blue arrowhead: autophagosome (n = 3). (C) Representative pictures of autophagosome formation monitored by MDC staining. Cell autophagy was analyzed by flow cytometry (n = 3). (D) Representative Western blots showing the protein levels of LC3II and SQSTM1 in adipocytes treated with leptin or TM followed by treatment with 400 nM bafilomycin A1 (BafA1), which was added in the last 4 h of the treatment period (n = 3). Full scans of uncropped blots are included in Figure S3. Values are means ± SEM. *p < 0.05 compared with the control group, # p < 0.05 compared with the TM group, & p < 0.05 compared with the BafA1 group.