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. 2007 Apr 12;26(10):2501–2512. doi: 10.1038/sj.emboj.7601685

Figure 4.

Figure 4

Retention of D18G TTR and amyloidogenic monomeric TTRs (M-TTRs) upregulates ER chaperone, BiP and GRP94. (A, E) HeLa cells were transfected with empty vector (E.V.) or the indicated M-TTR constructs. Cell lysates were recovered 48 h post-transfection, boiled and subjected to SDS–PAGE under reduced condition. Proteins were probed with anti-human TTR, anti-KDEL (BiP and GRP94), anti-PDI and anti-actin antibodies. As positive control for ER chaperones induction, cells transfected with empty vector were treated for 16 h with 2 μg/ml tunicamycin (Tm) 32 h post-transfection (Figure 4E, lane 2). (B–D and F–H) Densitometric quantifications were performed on BiP, GRP94 and PDI Western blots of HeLa cells. (B–D) Bars shown are the average expression of ER chaperones in cells transfected with non-amyloidogenic and amyloidogenic M-TTRs. (F–H) Bars shown are the average of three independent experiments for the expression ratio of ER chaperones in cells transfected with the indicated constructs. Values represent the mean±s.e. (**P<0.01, ***P<0.001)