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. 2018 Apr 10;26(3):431–444. doi: 10.3727/096504017X15031557924123

Figure 6.

Figure 6

Effect of GCNT3 on MCAM stability. (A) WM-115 cells were transiently expressed with aberrant GCNT3 or GFP as a control for 24 h and then treated with cycloheximide (final concentration of 10 μM). Following the indicated time intervals, cells were collected and analyzed for their intrinsic MCAM protein levels (left). The results from three independent experiments using the same protocols for MCAM are displayed by declined curves after normalization of the MCAM bands using their matched tubulin bands and subsequently adjusting the starting points to be at 0 h and 1.0 as standards (right). (B) Proposed pathway of GCNT3-mediated glycosylation is shown schematically. (C) GlcNAc levels of MCAM protein from the prepared cell samples were determined by immunoprecipitation and following detection with HRP-conjugated wheat germ agglutinin (WGA). WM-115 cells were transiently transfected or not transfected (−) with the indicated plasmids, GFP and GCNT3-Myc. WM-266-4 cells were treated or not treated (−) with the indicated siRNAs or Tal.