Figure S1.
Knockdown of AQP4 Suppresses Acute Cytotoxic Edema and Improves Functional Recovery in Rats at 3 dpi but Not at 28 dpi following DC Crush Injury
a, ELISA for AQP4 showing no increase in total AQP4 protein in primary astrocytes after 6 h under hypoxia (5% O2; n = 6); b, Representative immunoblot and c, densitometry to confirm 60% and 75% knockdown of AQP4 using in vivo JetPEI-delivered shRNA to AQP4 (shAQP4) at 3 dpi and 28 dpi following DC crush injury (n = 3 independent repeats from 3 pooled rat spinal cords/experiment (total n = 9 rats/condition)); d, Spinal cord water content was significantly suppressed at 3 dpi, but at 28 dpi the water content in DC + shAQP4-treated animals was significantly higher than DC + Vehicle-treated controls, despite 75% AQP4 protein knockdown (n = 3-4 rats/condition, 3 independent repeats (total n = 10 rats/condition)); e, Knockdown of AQP4 improved tape sensing and removal time up to 1 week after DC crush injury, but the sensing and removal time gradually increased to above DC + vehicle-treated controls at 4 weeks after DC crush injury (n = 3-4 rats/condition, 3 independent repeats (total n = 10 rats/condition)); f, The early improvement in ladder crossing ability of rats gradually worsened 1 week after and at 28dpi is higher than DC+vehicle-treated controls (n = 3-4 rats/condition, 3 independent repeats (total n = 10 rats/condition)). ∗ represents p < 0.05, ns represents p > 0.05 (see Table S2 for p values). Animals were euthanized at 28dpi due to episodes of vomiting, gait problems, lethargy and convulsions, possibly reflecting their inability to regulate water in the CNS. Related to Figure 2.