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. 2021 May 5;41(18):3948–3957. doi: 10.1523/JNEUROSCI.3019-20.2021

Figure 7.

Figure 7.

Expression of PKC98ECAAX in PKC98E conditional mutants restores sensitivity and ORCO S289 phosphorylation. A–C, Examples of anti-phospho-ORCO on (A) wild-type control (w1118), (B) PKC98E FRT conditional mutants, (C) PKC98E FRT conditional mutants expressing PKC98ECAAX (pORCO-GAL4, UAS PKC98ECAAX; PKC98E FRT, UAS-FLP). D, Quantitation of olfactory neuron dendrite fluorescence using phospho-S289 antiserum for the three genotypes; n = 5–10; ***p < 0.01 (p = 1.23 × 10−5 for PKC98E FRT conditional and w1118 control); +++p < 0.01 (p = 4.54 × 10−5 for PKC98E FRT conditional and PKC98ECAAX rescue); p values were not significant between w1118 and PKC98ECAAX (p = 0.396). E, Sample traces for w1118, PKC98E FRT conditional, and PKC98E FRT conditional mutants expressing PKC98ECAAX to 1% cVA. F, Dose–response curve to cVA for w1118, PKC98E FRT conditional, and PKC98E FRT conditional expressing PKC98ECAAX; n = 5–10. The sigmoidal curve for Δ spikes was plotted for different concentration of cVA with Hill fitting for the genotypes described; **p < 0.05, ***p < 0.01 (one-way ANOVA p values between PKC98E FRT conditional and w1118 for 0.01%, 0.03%, 0.1%, 0.3%, 1%, 3%, 10%, 30%, and 100% cVA are 0.530, 0.038, 0.011, 0.802, 0.003, 5.17 × 10−5, 0.002, 0.009, and 0.017, respectively). ANOVA comparison between PKC98E FRT conditional and PKC98ECAAX for the same cVA dilutions are 0.950, 0.090, 0.007, 0.214, 0.001, 0.004, 0.009, 0.007 and 0.011, and between PKC98ECAAX and w1118 are all below significance (p = 0.445, 0.441, 0.896, 0.422, 0.345, 0.163, 0.128, 0.260, and 0.618 for cVA doses, respectively). G, H, Dominant PKC98E flies are defective in desensitization. G, Representative responses of w1118 and PKC98ECAAX (pORCO-GAL4, UAS-PKC98ECAAX; PKC98E FRT, UAS-FLP) to 1% cVA with and without cVA preexposure. H, Desensitization of w1118 (left) and PKC98ECAAX (right). Black bars represent without preexposure to cVA. Gray bars represent responses after a 1-h cVA exposure. Preexposure significantly reduces cVA responses of w1118 (p = 0.001, two-tailed Student's t test, n = 10), but does not significantly affect PKC98ECAAX (p = 0.101, two-tailed Student's t test, n = 10). PKC98ECAAX shows impaired response reduction (interaction F(1,36) = 4.605, p = 0.039, two-way ANOVA); **p < 0.01. Data are mean ± SEM, ns, not significant.