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Fig. 4 Impact of inhibiting autophagy on the neuroprotective activities of RDM in SH-SY5Y cells treated with 6-OHDA. (A) Experimental flowchart. On day 1, cells were plated. On day 2, cells were pretreated with RDM for 1 h followed by treatment with 3-methyladenine (3-MA), an autophagy inhibitor. The cells were then subjected to 6-OHDA-induced neurotoxicity for 16 h. Cell viability was assessed using alamarBlue staining. (B) 3-MA cytotoxicity on SH-SY5Y cells using alamarBlue assay. Cells were treated with 3-MA at concentrations ranging from 1 to 100 μM for 16 h. The relative viability of control cells is presented as 100% for normalization. (C) Assessment of neuroprotective activity with 3-MA (10 μM) and RDM (10 μM) treatment. The relative neuroprotection (relative protection) was calculated based on cell viability, with the relative cell protection of untreated cells presented as 0%. *p < 0.05 compared with control group; #p < 0.05 compared with 6-OHDA alone group. RDM, rhodoptilometrin.