Figure S5.
rde-4 Mutants Are Defective in Chemotaxis under a Stressful Temperature but Display Normal Activity in the AWC Sensory Neuron, Related to Figure 6
(A and B) Results for experiments testing chemotaxis at day 1 of adulthood of worms. Chemotaxis index = ((# worms at stimulus)-(# worms at control)) / ((# total worms on plate)-(# worms at origin)). Each dot represents one plate with > 200 worms. All groups were tested on at least three independent trials (n = > 9).
(A) Chemotaxis screens were performed on RNAi factor mutants at both 20 (blue dots) and 25 degrees (red dots). Chemotaxis indices (x axis) were tested for the strains (y axis) N2(wild-type), rde-4(ne299), dcr-1(mg375), ergo-1 (tm1860), nrde-3 (gg66), rrf-1(ok589), rrf-3(pk1426), mut-16(pk710) and eri-6(mg379). The odor stimulus used was benzeldahyde (1:100). P values were determined by two-way ANOVA, ∗∗∗∗-p < 10−4.
(B) rde-4 mutants are defective in chemotaxis to multiple stimuli at high temperature. N2(wild-type) (black dots) and rde-4 mutants (green dots) raised at 25 degrees for chemotaxis to Benzaldehyde (1:102), Butanone (1:104), Diacetyl (1:103) and NaCl (50mM). P values were determined by Two-way ANOVA with Sidak’s post hoc correction for multiple comparisons. ∗∗∗∗- p < 10−4.
(C) A Principal Component Analysis (PCA) projection of 12 samples based on normalized STGs read counts. Each symbol represents one independent replicate. The corresponding genotype and temperature are indicated. The % variances, out of the total original variance in the high-dimensional space, spanned by the first and second Principal Components are indicated on the x- and y- axis, respectively. Related to Figure 6.
(D) Activity of sensory neuron AWC was quantified by GCaMP2 fluorescence intensity in a microfluidic device controlling stimulus exposure. N2 (wild-type) (n = 15) and rde-4 (n = 20) mutants were loaded into chips and exposed to the stimulus Isoamyl alcohol (1:104) for 1 minute followed by a switch to buffer (indicated by the red arrow). Each row represents an individual worm. Shown are the fluorescence intensity values normalized from 0 to 1 (Ft-Fmin)/(Fmax-Fmin) across time (seconds).
(E) Maximum fluorescence intensity increase (x axis) of the N2 and rde-4 worms (y axis) of AWCneurons in response to odor removal. Mean peak was defined as ΔF (2 s pre-stimuli) – max ΔF (post-stimuli).
(F) Time (x axis) it took from the moment of odor removal for AWC neurons in N2 and rde-4 worms (y axis) to reach maximum fluorescence intensity. P values were determined by Mann-Whitney tests. n.s.- p > 0.05.