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Research Paper Volume 10, Issue 8 pp 2190-2208
Qualitative and quantitative alterations in intracellular and membrane glycoproteins maintain the balance between cellular senescence and human aging
Relevance score: 12.268469Yoko Itakura, Norihiko Sasaki, Masashi Toyoda
Keywords: cellular senescence, human aging, intracellular glycan, lectin microarray, glycan profile
Published in Aging on August 29, 2018
Lectin microarray analysis for glycoproteins of TIG-3S, TIG-101, and TIG-102 at various PDLs. Schematic illustration of lectin microarray analysis for intracellular glycoproteins (enclosed by dotted line). Collected cells were heated and centrifuged with lysis buffer, containing 0.1% protease inhibitor, followed by fractionating as intracellular glycoproteins (upper layer) and membrane glycoproteins (lower layer).
Lectin microarray analysis for glycoproteins of TIG-3S, TIG-101, and TIG-102 at various PDLs. Heat map of log10-transformed lectin microarray data for intracellular glycans of TIG-3S, TIG-101, and TIG-102 compared to overall lectin-binding profiles at each PDL. Rows show 45 lectins and columns show PDLs of TIG-3S, TIG-101, and TIG-102 (27–94, 40–51, and 40–52, respectively). Lectin microarray data at each PDL were obtained from triplicate measurements. The color scale indicates low (green) to high (red) ratio. Underlines for lectins are shown as specific characters. Signature indicates lectin-binding types beside lectin names (closed circle; N-acetylglucosamine-oligomer; open circle, high-mannose; closed square, α2-6sialic acid; open square, α1-6fucose; closed triangle, galactose or high-mannose; open triangle, mannose- or complex-type N-glycan).
Biplot for PCA analysis of lectin microarray data in TIG-3S, TIG-101, and TIG-102. PC1 represents human aging. Pink, light blue, and dark blue labels represent TIG-3S, TIG-101, and TIG-102 cell lines, respectively. Color gradients (light to dark) of dots and numerals reflect progressive senescence in PDLs (i.e., young to aged). Left panel: cell passage replications, right panel: lectin replications.
Altered ratios of each lectin in intracellular glycans with cellular senescence. Line graphs show differences between lectin signal intensities at various PDLs and those at the first PDL in TIG-3S, TIG-101, and TIG-102. Changes in ratio were calculated based on average signal intensity at each PDL. Highest and the lowest values of the largest change in ratio are shown for each cell line. Each lectin is shown as a different color in a box.
Abundance of intracellular to cell surface glycans in seven lectins: SNA (A), SSA (B), ACG (C), MAH (D) at different PDLs. Left, middle, and right panels show TIG-3S, TIG-101, and TIG-102, respectively. Closed and open bars represent proportions of intracellular and cell surface glycans, respectively, relative to the total array signal at each PDL. Levels of the seven selected lectins in cell surface glycans changed with aging [10].
Abundance of intracellular to cell surface glycans in seven lectins: ECA (E), PWM (F), and WFA (G) at different PDLs . Left, middle, and right panels show TIG-3S, TIG-101, and TIG-102, respectively. Closed and open bars represent proportions of intracellular and cell surface glycans, respectively, relative to the total array signal at each PDL. Levels of the seven selected lectins in cell surface glycans changed with aging [10].
Localization of sialylated glycoproteins in TIG cells. (A) TIG-3S (PDL 38; top) and TIG-102 (PDL 46; bottom) stained with SNA (red; left panel), FITC-conjugated membrane marker (CD44, green; middle panel) and the overlay image (right panel). (B) TIG-3S (PDL 52; left) and TIG-102 (PDL 46; right) stained with FITC-conjugated SNA (green; left panel), an intracellular marker (mitochondria, red; middle panel), and the overlay image (right panel). Blue staining represents the nucleus.
Lectin blot detection of intracellular and membrane glycoproteins of TIG cells. The intracellular extracts from TIG-3S and TIG-102 and the corresponding membrane extracts were applied to lanes 1, 3 and 2, 4, respectively. They were subjected to lectin blot analysis using (A) biotinylated-SNA, (B) -MAH, (C) -WFA, and (D) -ECA.