Extended Data Fig. 7: MM-JH-1 incorporation in MGAT1 knockout HEK 293S GnTI- cells and MGAT1 mutated Lec1CHO cells.
From: Selective bioorthogonal probe for N-glycan hybrid structures
Cell lysates collected from HEK 293S GnTI- or Lec1CHO cells incubated with different concentrations of MM-JH-1 were subjected to click reactions with TRITC-azide to check the extent of cellular uptake. No incorporation of MM-JH-1 was observed in HEK 293S GnTI⁻ cells collected by (a) physical scraping. N = 3 independent biological replicates. (b) trypsinization. N = 3 independent biological replicates. (c) MM-JH-1 incorporated into HEK 293 T cells in a concentration dependent manner, but labeling was not detected in HEK 293S GnTI- cells. N = 3 independent biological replicates. No incorporation of MM-JH-1 was observed in Lec1CHO cells collected by (d) physical scraping. N = 3 independent biological replicates. (e) trypsinization. N = 3 independent biological replicates. (f) MGAT1 cDNA rescue on Lec1CHO cells resulted in a return in signal for both MGAT1 (detected with anti-GFP antibody) and MM-JH-1 labeling (detected with anti TRITC antibody) on immunoblot. N = 4 independent biological replicates.
