Translation visualization in neuronal somas. (A) Neuronal somas: from left to right: vehicle, puromycin, and HHT at first image cycle after treatment (top images) and tenth cycle (bottom images) after treatment was added. Red arrows point to puncta. (B–E) Graphs of number of puncta in V4-expressing cells (B,C) or V4-Arc expressing cells (D,E) normalized to first cycle after treatment (V4; vehicle n = 13 cells, puro n = 7 cells, HHT n = 12 cells; V4-Arc vehicle n = 14 cells, puro n = 8 cells, HHT n = 9 cells. Puncta were counted with an automated program (Materials and Methods). Experiments came from a minimum of three independent cultures. Vehicle groups from HHT (water) and puromycin (0.0675% Ethanol) were not significantly different from each other and were pooled. The same pooled vehicle groups are used in B–C and D–E. An interaction of time and group was observed for V4 (two-way ANOVA F(18,243) = 4.8, P < 0.0001) and V4-Arc (two-way ANOVA F(18,234) = 5.6, P < 0.0001). For V4, control vs. HHT and control vs. Puro, Tukey's post-hoc tests P < 0.0001 and <0.0001, respectively, and for V4-Arc control vs. HHT and control vs. Puro, Tukey's post-hoc tests P < 0.0001 and <0.0001, respectively. (F) Number of puncta at tenth image cycle (∼12 min after first image cycle) normalized to first image cycle immediately after treatment (n is same as B–E) measured using manual counting of puncta. Number of puncta was significantly different between controls and cells treated with HHT or puromycin for both V4 (one-way ANOVA F(31,2) P < 0.001), Tukey's post-hoc test P < 0.01 and V4-Arc (one-way ANOVA F(30,2) P < 0.001), Tukey's post-hoc test P < 0.01. All errors are shown as standard error of the mean. Asterisks are applied as follows: (**) P ≤ 0.01, (****) P ≤ 0.0001.
