Skip to main content
. 2018 Aug 16;14(11):1943–1958. doi: 10.1080/15548627.2018.1493043

Figure 2.

Figure 2.

6-OHDA treatment enhanced secretion of PARK7 from SH-SY5Y cells. (A and B) SH-SY5Y cells were treated with 0–100 μM 6-OHDA for 3 h and were then cultured in serum-free medium for 2 h. (A) Whole cell lysates and the conditioned medium were immunoblotted using antibodies specific for PARK7, RPN1, or ACTB. (B) PARK7 band intensities were quantified by densitometric scanning and the percentage of secreted PARK7/total PARK7 is shown. LDH release in the conditioned medium was analyzed by LDH assay. n = 3; mean ± S.D.; *, p < 0.05. (C) SH-SY5Y cells were pre-treated with 2 μg/ml brefeldin A for 3 h and were then treated with 100 μM 6-OHDA for 3 h, followed by culture in serum-free medium for 2 h. Whole cell lysates and the conditioned medium were immunoblotted with antibodies specific for PARK7, FN1, or RPN1. PARK7 and FN1 band intensities were quantified by densitometric scanning and relative secretion level to vehicle-treated cells is shown. n = 3; mean ± S.D.; **, p < 0.01; n.s., not significant. (D) SH-SY5Y cells were pre-treated with or without 20 μM pan-caspase inhibitor (zVAD) or CASP1 inhibitor (YVAD) for 1 h and were then treated with 100 μM 6-OHDA for 3 h, followed by culture in serum-free medium for 2 h. Whole cell lysates and the conditioned medium were immunoblotted using antibodies specific for PARK7 or RPN1. PARK7 band intensities were quantified by densitometric scanning and relative secretion level to vehicle-treated cells is shown. n = 3; mean ± S.D.; **, p < 0.01.