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. 2018 Dec 21;8(1):bio039511. doi: 10.1242/bio.039511

Fig. 3.

Fig. 3.

Effect of membrane cholesterol depletion by MβC on neuropathic manifestation in RTX neuropathy. (A,B) Effect of membrane cholesterol depletion in the mechanical allodynia of RTX neuropathy. Membrane integrity was disrupted by four doses of MβC (1 µg/day, cumulative 4 µg per mouse) through an intrathecal lumbar puncture, and the changes in mechanical threshold were evaluated at day 7 of RTX neuropathy (RTXd7), RTXd14 and RTXd21 as described in the Materials and Methods. Cholesterol depletion by MβC delivered before RTX neuropathy (MβC pre-RTX group, A) delayed the onset of mechanical allodynia, but MβC had no effect on mice in which RTX neuropathy was established (MβC post-RTX, B). For MβC pre-RTX, MβC was delivered at RTXd0, RTXd1, RTXd3 and RTXd5; for Sal+RTX, animals with RTX neuropathy received additional saline as control; for MβC alone, naïve mice received MβC as per the schedule of the MβC pre-RTX group; for the MβC post-RTX, MβC was delivered at RTXd7, RTXd9, RTXd11 and RTXd13. Arrows indicate the time points of RTX administration. *P<0.05: pairing repeated measures analysis of variance (ANOVA) followed Tukey post hoc test comparing before and after of MβC administration in the MβC pre-RTX group. #P<0.05: nonpairing repeated measures ANOVA followed Tukey post hoc test between the MβC pre-RTX and Sal+RTX groups. (C) Thermal responses after RTX neuropathy were evaluated using the hot plate test at 52°C as described in the Materials and Methods. Thermal hypoalgesia was induced by RTX and MβC had no effect on the thermal responses. **P<0.01 Sal+RTX, MβC pre-RTX, and MβC post-RTX groups compared with the MβC alone group. (D) Total cholesterol content of the DRG tissues was evaluated using a cholesterol assay kit following the manufacturer's instructions in the vehicle group (open diamond, n=8) and at d7 (open square, n=7) and d21 (open circle, n=8) in the MβC alone group. This graph indicates that MβC depleted membrane cholesterol and resulted in disrupting the membrane integrity. ***P<0.001: d7 of the MβC alone group compared with the vehicle group. #P<0.05: d7 compared with d21 of the MβC alone group.