Supplementary Figure 3: GFP-PAC-1 transgene expression levels and controls for GFP-PAC-1N localization in the absence of endogenous PAC-1. | Nature Cell Biology

Supplementary Figure 3: GFP-PAC-1 transgene expression levels and controls for GFP-PAC-1N localization in the absence of endogenous PAC-1.

From: An instructive role for C. elegans E-cadherin in translating cell contact cues into cortical polarity

Supplementary Figure 3

(a) Expression levels of GFP-PAC-1 transgenes and derivatives, measured by fluorescence intensity (after background subtraction) at the four-cell stage (see Methods), expressed in arbitrary fluorescence units. PAC-1 amino acids present in each GFP fusion protein are indicated (ΔPH lacks the PH domain only). The box represents first and third quartiles, bars represent maximum and minimum values, and the line within the box is the mean fluorescence intensity (full length n = 11 embryos, ΔPH n = 13, 392-838 n = 11, 575-1604 n = 13, 2-610 n = 12, 1-574 n = 14). While expression levels vary between transgenes, contact localization is evident even from those with the lowest expression levels (ex. full-length GFP-PAC-1 and GFP-PAC-12–610). Therefore, lack of apparent contact localization for GFP-PAC-1392–838 and GFP-PAC-1575–1604 is not due to low expression level. (b) Full-length GFP-PAC-1 in control (empty vector) RNAi embryos localizes to cell contacts (arrow, 20/20 embryos). (c) In pac-1(3′ RNAi) embryos GFP-PAC-1 is not detected (19/19 embryos), indicating that RNAi effectively depletes PAC-1. Scale bars, 10 μm.

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