Fig. 2

Metabolic characterization of CD151 depletion in osteosarcoma. a Liquid chromatography coupled to mass spectrometry (LC/MS) was used to measure the concentration of intermediates in HOS-MNNG cells. Heatmap showing significantly differently expressed metabolites altered by CD151 silencing. Shades of red and blue represent higher and lower levels of metabolites, respectively. b The top 10 enriched pathways from integrated pathway analysis of significantly changed metabolites. The red indicates the KEGG pathway related to sphingolipid metabolism. c Lipids were extracted from WT or CD151 KO HOS-MNNG cells and analyzed by LC/MS. Bubble plots represent the mean log₂-transformed fold-change difference between cell lines. d Representative images and quantification of BODIPY FL-labeled ceramide or sphingomyelin was analyzed by confocal fluorescence imaging in HOS-MNNG cells. Scale bars: 50 μm. e HOS-MNNG cells with CD151 depletion were incubated with Alexa 555-conjugated CTB (red) to label GM1-containing lipid rafts. Cells were analyzed by confocal microscopy or flow cytometry. Scale bars: 50 μm. Comparisons were made using the two-tailed, unpaired Student’s t-test; *p < 0.05, **p < 0.01, ***p < 0.001