Fig. 7.

Chronic treatment with fluoxetine induces GluK2 editing in cultured astrocytes, which requires ADAR2 activity. Cells were treated with transfection solution without siRNA (siRNA (−); open symbols) or with siRNA specific to ADAR2 (siRNA (+); solid symbols). Three days later, cells were either not further treated (triangles) or treated with 10 μM of fluoxetine (diamonds) for 3 days. Thereafter, cells were collected for amplification refractory mutation system polymerase chain reaction (ARMS-PCR) analysis of GluK2 editing at (A) I/V, (B) Y/C and (C) Q/R sites. Representative Southern blots showing edited (guanosine; G) and unedited (adenosine; A) PCR products at (A1) I/V, (B1) Y/C or (C1) Q/R sites. Similar results were obtained from 3 independent experiments. Scans of known mixtures of G and A (n = 3) were made to prepare calibration graphs showing scanning intensity (as a fraction of maximum intensity) as a function of G/(G+A) for each mixture (A2, B2 and C2; solid circles). Based on the calibration curves and scans of G/(A+G) (n = 3) under each condition the percentages of edited (G) GluK2 at each site was determined (A2, B2 and C2). Standard error of the mean values are indicated by horizontal bars. *Statistically significant (p < 0.05) difference from all other groups.