Fig. S1. Immunologic characterization of endogenous EGL-4. (A) Western blot of 100 μl of either wild-type (N2, first two lanes) or
egl-4(
n479) mutant animals boiled for 30 min in Lamelli sample buffer and resolved on an 8% polyacrylamide gel with SDS. The first lane was incubated with preimmune serum, and the subsequent lanes were incubated with serum from a rabbit that had been exposed to a fragment of EGL-4. The left arrow indicates the mobility of full-length EGL-4, which is close to its predicted molecular weight of 87 kDa. (B-D) Fluorescence micrographs of the anterior ganglion of a wild-type animal that expressed GFP under the
str-2 promoter to outline the AWC neuron are shown. The arrowhead points to the AWC neuron, the red line in C outlines the cell body, and the white indicates immunoreactivity with the same antiserum used in A above. Similarly treated
egl-4(
n479) animals showed no staining using this serum and these procedures (1). (E) Benzaldehyde exposure results in nuclear endogenous EGL-4. Animals were exposed to buffer (Upper) or benzaldehyde (Lower) for 80 min before fixing and staining (2), anti-EGL-4 antibodies were followed by DAPI and rhodamine-conjugated secondary antibodies. The AWC cell body was visualized by
pstr-2::GFP and outlined in green, whereas the nucleus, outlined in red, was visualized by DAPI. The staining seen in E is rhodamine. All eight benzaldehyde-exposed animals exhibited nuclear EGL-4 staining as seen in Upper, whereas 10 of 12 naive animals showed the cytoplasmic EGL-4 seen in Lower.