Figure 2.

Dose–response stimulatory effects of wild‐type and mutant NGF on proliferation and differentiation in primary human foetal dorsal root ganglion (DRG) neurons. Immunofluorescent automatic high‐content image analysis was performed as described in Materials and Methods2. Human foetal DRG neurons were treated with 10‐fold serial dilutions of wild‐type NGF (black circles ●), NGF‐R100E (red squares ), NGF‐W99A (green diamonds ) and NGF‐K95A/Q96A (blue triangles ) on day 0 and incubated for 4 days. Dose‐dependent effects of wild‐type or mutant NGF on (a) proliferation, as judged by Hoechst and β‐tubulin‐positive cells, or (b) differentiation, as judged by average neurite length per cell. (c) Normalized dose‐dependent effects of wild‐type NGF or NGF‐K95A/Q96A on differentiation at high concentrations. Three independent biological repeats with eight (a and b) or four (c) technical replicates were conducted