(A) PC12 cells were transfected with indicated siRNAs for 48 hours and then treated with 50 ng/ml recombinant NGF. Images were taken at the indicated time points (left). Percentage of differentiated cells was quantified (right). Scale bar, 20 μM. Bars, mean±SD biological triplicate of experiments.
(B) PC12 cells transfected with indicated siRNAs (left) or expressing indicated constructs (right) were seeded in 96 well plates at a density of 2,000 cells/well and supplemented with or without 100 ng/ml EGF. Cell growth was measured. Bars, mean±SD biological triplicate of experiments. **** p<0.0001, two-tailed Student’s t test.
(C) PC12 cells expressing doxycycline-inducible A-, B- or C-RAF were treated with 100 ng/ml doxycycline for 24 hours. Cells were then treated with 100 ng/ml EGF. Scale bar, 20 μM. Bars, mean±SD biological triplicate of experiments.