Fig. 5: Regulation of NPC phenotypes on GMOC through MFG-E8 enrichment.
From: An enzyme-proof glycan glue for extracellular matrix to ameliorate intervertebral disc degeneration
MA plot for different proteins between GMOC and (A) glass or (B) GMAC. C Heatmap of differentially enriched membrane proteins of NPCs between glass, GMAC, and GMOC, as determined by 4D label-free quantitative proteomic analysis. IF staining and quantification for (D, E) MFG-E8 (red) and (F, G) Integrin alpha-V (green) in NPCs cultured on GM substrates or glass, with nuclei counterstained with DAPI (blue) (scar bar: 200 μm; Data are presented as mean values ± SD, n = 5, biological replicates). H The representative image showing live/dead staining of NPCs on GM esters with or without Mfge8 siRNA treatment. Green: living cell; red: dead cell (scar bar: 400 μm). I The representative image showing IF staining for MFG-E8 in NPCs on GM esters with or without Mfge8 siRNA treatment. MFG-E8 (red) and nuclei (DAPI, blue) (scar bar: 100 μm). J IF staining for Type II collagen in NPCs on GM esters with or without Mfge8 siRNA treatment. Type II collagen (red), Cytoskeleton (phalloidin, green) and nuclei (DAPI, blue) (scar bar: 50 μm, n = 5, biological replicates). K Quantitative analysis of Type II collagen IF staining (Data are presented as mean values ± SD, n = 5, biological replicates). L Dot-blotting analysis of secreted MFG-E8 from living NPCs on GM substrates, with or without Mfge8 siRNA treatment (Data are presented as mean values ± SD, n = 4, biological replicates). M The representative image showing IF staining for cytoskeleton (phalloidin, green) and nuclei (DAPI, blue) in NPCs on GM esters, with or without Integrin αv antagonist MK0429 treatment (scar bar: 200 μm). N Western Blotting analysis of Akt and p-Akt and (O) quantification of gray value in NPCs cultured on the GMOC. All samples derive from the same experiment, and blots were processed in parallel. (Data are presented as mean values ± SD, n = 5, biological replicates). Data were analyzed using the Kruskal-Wallis test, followed by Dunn’s post hoc test for multiple comparisons (E, G), or two-sided Mann-Whitney test (K, L, O). Source data is provided as a Source Data file.
