Journal: Biogerontology

Article Title: Glycan characteristics of human heart constituent cells maintaining organ function: relatively stable glycan profiles in cellular senescence

doi: 10.1007/s10522-021-09940-z

Figure Lengend Snippet: Lectin blot detection of whole cell and membrane extracts from heart constituent cells. a Whole cell extracts from HCMs, HCFs, HCAECs, and HMVECs were applied to lanes 1–4, respectively and were subjected to lectin blot analysis using biotinylated-SNA, -WFA, -RCA120, -TJA-II, and -AOL. b The membrane extracts from HCMs, HCFs, and HCAECs at T1 (growth phase) and the corresponding extracts at T2 (growth arrest) were applied to lanes 1, 3, 5 and 2, 4, 6, respectively and were subjected to lectin blot analysis using biotinylated-SNA, -RCA120, -PHA-E, -WFA, -AAL, and -AOL. Bar graph representing the bands for each lane after normalization to HCMs using the Fusion system. All experiments were performed with repetition. The data are presented as the mean ± SD (n = 3)

Article Snippet: After blocking, the membrane was washed with Tris-buffered saline containing 1% Tween 20 (TBST) and reacted with each of 1 µg/mL biotinylated SNA (EY Laboratories Inc., San Mateo, CA, USA), 2 µg/mL biotinylated WFA (EY Laboratories Inc.), 1 µg/mL biotinylated RCA120 (Vector Laboratories Inc., Burlingame, CA, USA), 1 µg/mL biotinylated AAL (J-OIL MILLS Inc., Tokyo, Japan), 2 µg/mL biotinylated TJA-II (Seikagaku Biobusiness, Tokyo, Japan), 1 µg/mL biotinylated AOL (Tokyokasei Industry Co., LTD., Tokyo, Japan), and 1 µg/mL biotinylated PHA-E (EY Laboratories Inc.) at 25 °C for 1 h. Before use, TJA-II and AOL were biotinylated using a Biotin Labeling Kit-NH 2 (DOJINDO, Kumamoto, Japan).

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