FAM134B-Mediated ER-phagy Upregulation Attenuates AGEs-Induced Apoptosis and Senescence in Human Nucleus Pulposus Cells

Figure 3

AGEs treatment upregulated FAM134B-mediated ER-phagy in human NP cells. (a–d) The human NP cells were exposed to 200 μg/mL AGEs for different times (0, 6, 12, 24, and 36 h); ER-phagy-associated protein levels of FAM134B, LC3, and p62 were detected by western blot assay, and relative band density was quantified. (e, f) After treated with 200 μg/mL AGEs for 36 h, relative protein expression of FAM134B and LC3 were assessed using immunofluorescence staining; representative images and relative intensity quantification were illustrated, scale bar: 50 μm. (g) ER and lysosome colocalization profile was detected by ER-tracker and Lyso-tracker staining. (h, i) Transmission electron microscopy results for ER positive autophagosomes/autolysosomes (as indicated by black arrow), scale bar: 1 μm and 500 nm. Data are represented as . , .

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)