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Fig. 3. Reduced AQP4 expression, water permeability and migration in AQP4 RNAi-treated wild-type astroglia. (A) Phase-contrast micrographs of wild-type astroglial cell cultures after exposure to AQP9 (control) RNAi (left) or AQP4 RNAi (right). (B) Immunoblot of AQP4 expression in wild-type astroglia (+/+) and AQP4-null astroglia (–/–) without pre-treatment (blank), or with pre-treatment (arrow) with AQP4 RNAi, AQP9 RNAi or oligofectamine (+ Oligo). Positions of molecular size markers in kDa are indicated. (C) Osmotic water permeability of astroglial cells with and without AQP4 RNAi treatment. Changes in calcein fluorescence signal (left) in response to hypo-osmolar medium (150 mOsm) and return to iso-osmolarity (300 mOsm). Summary (right) of relative rates of water transport (mean±s.e.m.; n=3-4 cultures/condition; *P<0.01). (D) Summary of Transwell migration data for 1% or 10% serum, labeled as in B. Data are the mean±s.e.m. The number of Transwells used for migration is shown over each data bar in parentheses. *P<0.05 compared with migration of untreated wild-type astroglial cells. Bar, 200 µm.

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