Fig. 5. The analysis of tau protein under the modulation of Bmal1 in vitro. (A) The qRT-PCR (upper) and western blot (lower) data show that Bmal1 overexpression increases the mRNA and protein levels of Hsp70 in HEK293T cells. Data are shown in mean±SEM (n=3). *p<0.05 on Mann-Whitney test. (B) The marked reduction of tau levels is noted in all tested fractions after Bmal1 overexpression. Data are mean±SEM (n=3). *p<0.05 and **p<0.01. (C) The transfection of the short-hairpin RNA vector for Hsp70 (shHsp70) effectively diminishes the transcription (left) and protein expression (right) of Hsp70. The diminution of tau protein levels by Bmal1 overexpression is attenuated by shHsp70 (arrows) when it is compared with the short hairpin RNA empty vector (shControl). (D) The treatment of CQ (5 μM) or MG132 (2.5 μM) attenuates the Bmal1-mediated tau decrease as shown by an increase of tau protein levels in the presence of CQ and MG132 on western blot. EV, empty vector; FC, fold-change; sh, short hairpin.
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