Fig 3.
slo-1 and
slo-2 are expressed asymmetrically in the AWCON neuron. (A, C) Images of wild-type L1 animals showing expression of
slo-1p::GFP (A)and
slo-2p::GFP (C) at a higher level in AWCR (bottom panels) than in AWCL (top panels). Both AWCL and AWCR were labeled by
odr-1p::TagRFP. Thecell body of both AWC cells is outlined by dashed lines. Scale bar, 5 um. Anterior is at left and ventral is at bottom. (B, D) Quantification of asymmetric expression of
slo-1p::GFP (B) and
slo-2p::GFP (D) in AWCL and AWCR in wild type and mutants defective in AWC asymmetry. The single focal plane with the brightest fluorescence in each AWC was selected from the acquired image stack and compared for fluorescence intensity. The fluorescence intensity of
slo-1p::GFP and
slo-2p::GFP was compared using visual quantitative scoring between AWCL and AWCR in each animal, as previously performed [7,22,58].If no obvious difference in fluorescence intensity between the two AWC cells was observed, the animal was categorized as AWCL = AWCR. If an obvious difference in fluorescence intensity was observed between AWCL and AWCR, the animal was assigned to AWCL > AWCR or AWCR > AWCL. For both
slo1p::GFP and
slo-2p::GFP, the visual quantification of fluorescence was performed by the same individual. Only animals with visible expression in both AWC neurons were used in the analysis. p values were calculated using Fisher's exact test. ns, not significant. Error bars indicate standard error of proportion. (E, G) Representative images of wild-type L1 animals expressing
slo-1p::2xnlsGFP (E) and
slo-2p::GFP (G) in AWCON (bottom panel) but not in AWCOFF (top panel). Both AWC neurons were marked with
ceh-36p::myrTagRFP. AWCON cells were marked by
str-2p::2xnlsTagRFP, and AWCOFF neurons were defined by lack of
str-2p::2xnlsTagRFP. Scale bar, 5 um. Anterior is at left and ventral is at bottom. (F, H) Quantification of
slo-1p::2xnlsGFP (F) and
slo-2p::GFP (H) expression in AWCON and AWCOFF. The single focal plane with the brightest fluorescence in each AWC was selected from the acquired image stack and compared for fluorescence intensity. The fluorescence intensity of
slo-1p::2xnlsGFP and
slo-2p::GFP was compared using visual quantitative scoring between AWCON and AWCOFF in each animal, as previously performed [7,22,58]. Each animal was categorized into one of three categories: AWCON = AWCOFF, AWCON > AWCOFF, and AWCOFF > AWCON based on the comparison of GFP intensities between AWCON and AWCOFF cells of the same animal. p values were calcula