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. 2013 Jan 14;288(8):5357–5363. doi: 10.1074/jbc.C112.421768

FIGURE 3.

FIGURE 3.

Effect of N-linked glycosylation on the secretion of recombinant hLOXL2s and hLOX from S2 cells. A and B, Western blot analyses of the expression profiles of the glycosylation site mutants of Δ1-3SRCR-hLOXL2 (A)(nonadjacent lanes from the same blot are digitally juxtaposed) and Δ1-4SRCR-hLOXL2 (B) from induced transiently transfected cells. M, cell culture medium; S, soluble cell lysate; I, insoluble cell lysate (cell pellet). C, PCR confirmation of the stable integration of the N455Q and N644Q mutants of Δ1-3SRCR-hLOXL2 into the S2 genome. Stably integrated WT Δ1-3SRCR-hLOXL2 was used as a positive control. D and E, Western blot analyses of the expression profiles of unglycosylated WT Δ1-3SRCR-hLOXL2 (D) and WT Δ1-4SRCR-hLOXL2 (E) from tunicamycin-treated induced stable cells. Panels D and E are γ-adjusted to enhance minimally visible bands. Tunica., tunicamycin. F, Western blot analysis of expression of the LOX catalytic domain of hLOX in stable S2 cells.