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. 2017 Dec 15;9(5):5562–5577. doi: 10.18632/oncotarget.23726

Figure 1. Ganglioside GM1 levels increase in TNFα-treated HAECs.

Figure 1

(A) Levels of GM1 in TNFα-treated HAECs analyzed by flow cytometry. Representative results are shown. GM1 expression in control cells and negative controls is depicted by black dot lines and thin gray lines, respectively. (B) Mean fluorescence intensities (MFIs) of GM1 expression relative to control HAECs. (C) Representative photos of HAECs 3 days after exposure to TNFα (0.1 ng/ml–10 ng/ml). Inset image is at high magnification. (D) Cell surface levels of gangliosides in short-term (3 days) 1 ng/ml TNFα-treated HAECs analyzed by flow cytometry using antibodies against each ganglioside. Representative results are shown. Negative controls are depicted by thin gray lines. (E) MFIs relative to control HAECs. Results are presented as means ± SD from three independent experiments. (F) Pathways of gangliosides and glycosyltransferases contributing to each synthetic pathway. Gangliosides highlighted in red fonts were examined in this study. (G) Real-time PCR analysis of the glycosyltransferases shown in (F) and NEU3 using cDNA derived from control and short-term (3 days) 1 ng/ml TNFα-treated HAECs. The results are shown after normalization against values obtained for control HAECs (value = 1). Results are presented as means ± SD from three independent experiments. *P < 0.05; **P < 0.01. Control (Ctr): untreated cells.