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. 2021 Feb 18;18(8):1810–1823. doi: 10.7150/ijms.51654

Figure 4.

Figure 4

HSM regulates autophagosomes accumulation in macrophages dependent on TLR4 activity. (A) The effect of different concentrations HSM on the activity of RAW264.7 cells was analyzed by CCK-8 assay. (B) In vitro experiment to explore the effect of HSM on CQ-induced autophagy flux accumulation. RAW264.7 cells were pretreated with CQ (20 μM) for 2 h and then treated with or without HSM (16 μg/ml) for 16 h. Representative immunofluorescence image of LC3B in RAW264.7 cells treated with CQ and HSM. Scale bar: 40 μm. (C) The effect of HSM on the expression of LC3B mRNA was analyzed by q-PCR. (D) Western blotting to examine the effect of HSM on the expression of LC3B protein. (E) RAW264.7 cells were pretreated with CQ (20 μM) and CLI095 (100 μg/ml) for 2 h and then treated with or without HSM (16 μg/ml) for 16 h. Representative immunofluorescence image of LC3B in RAW264.7 cells treated with CLI095. Scale bar: 30 μm. Data are presented as means ± SEM of at least three separate experiments. *P < 0.05, **P < 0.01, ***P < 0.001.