Table 2.

The inhibitory effects of the Tiam1 antisense oligonucleotides are reversible

Antisense treatment	Hours after the release from the antisense treatment
Antisense treatment	0	12	24
Non-treated
Tiam fluorescence intensity	138 ± 14	135 ± 8	141 ± 11
Growth cone area	195 ± 15		206 ± 8
% Stage 2 neurons	68 ± 5		10 ± 2
% Stage 3 neurons	26 ± 3		85 ± 4
Sense treated
Tiam1 fluorescence intensity	148 ± 6		136 ± 14
Growth cone area	225 ± 20		215 ± 10
% Stage 2 neurons	67 ± 4		7 ± 4
% Stage 3 neurons	27 ± 6		89 ± 6
AST1a treated (2.5 μm)
Tiam1 fluorescence intensity	12 ± 2^2-a	38 ± 6^2-a	107 ± 10
Growth cone area	48 ± 11^2-a	95 ± 10	147 ± 14^2-a
% Stage 2 neurons	89 ± 5^2-a		42 ± 8^2-a
% Stage 3 neurons	4 ± 1^2-a		52 ± 5^2-a
AST1a treated (0.5 μm)
Tiam1 fluorescence intensity	55 ± 8^2-a		127 ± 8
Growth cone area	108 ± 9^2-a		185 ± 14
% Stage 2 neurons	75 ± 5		12 ± 4
% Stage 3 neurons	15 ± 5		79 ± 6

Cultures were treated with the oligonucleotides for 24 hr; after that time period the medium was replaced by a fresh one lacking oligonucleotides, and the cells were analyzed at different time intervals after the release of the treatment. Data are expressed as the mean ± SEM. Tiam1 levels were determined by quantitative fluorescence as described in Materials and Methods. Measurements were performed within the cell body and are expressed in pixels. Area values are expressed in micrometers squared. Measurements were performed in the largest growth cone that stage 2 or stage 3 neurons displayed. A total of 100 cells were analyzed for each time point and experimental condition. The percentage of stage 2 or 3 cells was measured in six different coverslips for each experimental condition. A total of 100 cells were analyzed in each coverslips.
↵F2-a Values significantly different from those of the control groups.