Fig. 6.

Fig. 6.

Multiplexed cellular expression tagging reveals vamp1/2 co-expression in motor axons. (A) Our optimized PCR tagging conditions facilitate multiplex knock-in in two target loci simultaneously. (B) Example memmScarlet-2A-vamp1⁺; memEGFP-2A-vamp2⁺ F0 larva shows partially overlapping expression patterns, e.g. in motor axons (arrowheads). (C) Efficiency of double cellular tagging of vamp1/2, shown as percentage of F0 larvae showing memEGFP and/or memmScarlet fluorescence. Bars represent one injection round, error bars indicate 95% confidence intervals for FP⁺ expression (Wilson/Brown calculation). (D) vamp1-mRuby3 or vamp2-mCherry knock-in lines crossed with transgenic motor neuron reporter mnx1:GFP confirms that both Vamp1 and Vamp2 proteins are found at the presynaptic terminals of motor neurons, i.e. the neuromuscular junctions that innervate the trunk musculature. Scale bars: 5 µm.