Figure 6.

Phenotyping of FtMt expressing neuronal cells. (A,B) Measurements of Cell Growth Rates of Undifferentiated FtMt clones. (A) Results showing relative increase in cell area occupied as percentage of Day 1 of FtMt clones and untransfected cells. Results show analyses at Day 7. Measurements involved area occupied of growing cells identified using Image J image analysis software. Data analyzed by Two-way ANOVA. Relative statistical differences between clones and are listed in Table 2A. (B) Representative photomicrographs of proliferating clones untransfected, Medium FtMt expressor and High B FtMt expressor at Day 1, Day 3, Day 5, and Day 7. (C,D) Measurements of Neurite formation of Differentiated FtMt clones. (C) Results showing relative area occupied of neuritic processes at day 7 of untransfected (Un), Medium FtMt expressor (Med) and High Ft expressor (High B). Measurements involved measuring area occupied of neurites identified using NeuronJ plugin of Image J image analysis software. Results are representative of triplicate experiments. Relative statistical differences between clones listed in Table 2B. (D) Representative photomicrographs of differentiated, untransfected, Medium FtMt expressor and High B FtMt expressor at Day 7. (E,F) Responses of FtMt overexpressing clones to oxidative stress. (E) Changes in cell viability (as percentage of untreated cultures) with increasing doses of hydrogen peroxide (H₂O₂) (0–250 μM). Untransfected and FtMt expressing clones were differentiated in microtiter plate wells for 7 days. Treatments added to media lacking RA containing 1% FBS on day 7 for 24 h. Cell viability assessed by added WST-1 reagent after 24 h. Absorbance (450 nm) measured after 1, 2, and 4 h. Results presented (mean ± SEM) of six wells/treatment and combination of three independent experiments. Data analyzed by Two Way ANOVA. Relative statistical differences between clones and treatments are listed in Table 3A. (F) Changes in cell viability (as percentage of untreated cultures) with increasing doses of Cobalt Chloride (CoCl₂) (0–200 μM). Untransfected and FtMt expressing clones were differentiated in microtiter plate wells for 7 days. Treatments added to media lacking RA containing 1% FBS on day 7 for 24 h. Cell viability assessed by added WST-1 reagent after 24 h. Absorbance (450 nm) measured after 1, 2, and 4 h. Results presented (mean ± SEM) of six wells/treatment and combination of three independent experiments. Data analyzed by Two-way ANOVA. Results indicate level of significance. *p < 0.05; **p < 0.01, ***p < 0.001, ****p < 0.0001. Relative statistical differences between clones and treatments are listed in Table 3B.