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. 2015 Sep 6;5:763–773. doi: 10.1016/j.fob.2015.09.002

Fig. 6.

Fig. 6

Involvement of GXXXG motifs in CTF dimerization and Aβ production. CHO cells were transfected with C99-hGLuc1 and 2 or C83-hGLuc1 and 2 and their GXXXG Flemish (Fle) and mutant 5 (mut5) corresponding mutants. (A) Cells transfected with the control empty vector (mock), the C99-hGLuc1 and 2, C99Fle-hGLuc1 and 2 or C99mut5-hGLuc1 and 2 proteins. Protein expression was monitored in cell lysates by Western blotting with the W0-2 or hGluc antibodies (top panels). Luciferase activity was measured and expressed as RLU normalized to non-mutated C99 (bottom). Values (means ± SEM) are representative of 5 independent experiments (n = 4 in each experiment). *p < 0.05, **p < 0.01 and ***p < 0.001, as compared to C99-hGLuc1 and 2. (B) Cells transfected with the control empty vector (mock), the C83-hGLuc1 and 2, C83Fle-hGLuc1 and 2 or C83mut5-hGLuc1 and 2 proteins. Protein expression was monitored in cell lysates by Western blotting with the Cter or hGLuc antibodies (top panels). Luciferase activity was measured and expressed as RLU normalized to non-mutated C83 (bottom). Values (means ± SEM) are representative of 3 independent experiments (n = 4 in each experiment). *p < 0.05, **p < 0.01 and n.s. (non significant), as compared to C83-hGLuc1 and 2. (C) Aβ 38, 40 and 42 production for C99-hGLuc1 and 2, C99Fle-hGLuc1 and 2 or C99mut5-hGLuc1 and 2 was measured by ECLIA in the culture media and given in pg/ml. Values (means ± SEM) are representative of 3 independent experiment (n = 4 in each experiment). ***p < 0.001, as compared to non-mutated C99.