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. 2020 Apr 27;10(13):5829–5844. doi: 10.7150/thno.44051

Figure 2.

Figure 2

Trehalose induces autophagy in cisplatin-treated HK2 cells. (A) The expression of the proteins P62 and LC3 was measured by western blotting in HK2 cells exposed to cisplatin (5 μM, 10 μM, 20 μM). (B) The expression of the proteins P62 and LC3 was measured by western blotting in HK2 cells exposed to cisplatin (5 μM) in the presence or absence of HCQ (20 μM). (C) HK2 cells were treated with cisplatin (5 μM) in the presence or absence of Rapa (50 nM) or HCQ (20 μM), and cell viability was determined. (D) The expression of the proteins P62 and LC3 was measured by western blotting in HK2 cells exposed to trehalose (20 mM, 50 mM, 100 mM). (E) The expression of the proteins P62 and LC3 was measured by western blotting in HK2 cells exposed to trehalose (50 mM) in the presence or absence of HCQ (20 μM). (F) HK2 cells treated with trehalose (50 mM) and p62 mRNA was measured by real-time polymerase chain reaction (real-time PCR). Data analyzed by Mann-Whitney. (G and H) HK2 cells were exposed to cisplatin (5 μM) in the presence or absence of trehalose (50 mM), and the expression of the proteins P62, LC3, Pink1 and Parkin was measured by western blotting. (I) Representative images of the colocalization between LC3 and mitochondria. Data are shown as the means ± s.d. from at least three independent experiments. A - C and D - I, Data analyzed by one-way ANOVA with Tukey's test. * P < 0.05, ** P < 0.01 vs. Con; & P < 0.05, && P < 0.01 vs. Cisp; # P < 0.05, ## P < 0.01 vs. HCQ. (Con, control; Cisp, cisplatin; HCQ, hydroxychloroquine; Rapa, Rapamycin; Tre, trehalose; Cisp + Tre, cisplatin + trehalose).