Supplementary Figure. 1: Mono-trans-synaptic spread of RV-Flp-CFP viruses depends on expression of RG in starter PNs (Related to Figs. 1 and 2). | Nature Neuroscience

Supplementary Figure. 1: Mono-trans-synaptic spread of RV-Flp-CFP viruses depends on expression of RG in starter PNs (Related to Figs. 1 and 2).

From: Intersectional monosynaptic tracing for dissecting subtype-specific organization of GABAergic interneuron inputs

Supplementary Figure. 1

(a) Experimental schema for monosynaptic tracing of input neurons sending direct inputs to supragranular PNs. (b) Confocal projection image merging H2BYFP (yellow) and CFP (cyan). CFP + /H2BYFP + and CFP + /H2BYFP- represent starter PNs, and general input neurons respectively of animals prepared as shown in a. Scale bar, 500 μm. (c,d) Confocal projection images merging DAPI (blue) and CFP (cyan) signals in the contralateral SSC (c) and the thalamus (d) of animals prepared as shown in a. Scale bar, 200 μm.(e) Quantification of the laminar distribution of CFP + /H2BYFP + starter PNs (e; L1: 0 %, L2/3: 89.9 ± 4.3 %, L4: 10.1 ± 4.3 %, L5: 0 %, L6: 0 %; 7680 cells, 3 animals) and CFP + /H2BYFP- general input neurons (f; L1: 0.2 ± 0.2 %, L2/3: 55.1 ± 2.9 %, L4: 28.7 ± 0.9 %, L5: 15.1 ± 1.9 %, L6: 1.0 ± 0.1 %; 23,317 cells, 3 animals)(g) Experimental schema for control experiments that test dependency of RV trans-synaptic spread on RG expression in starter PNs. (h) Confocal projection image merging H2BYFP (yellow) and CFP (cyan). CFP + /H2BYFP + and CFP + /H2BYFP- represent starter PNs, and general input neurons respectively of animals prepared as shown in g. Scale bar, 500 μm.(i,j) Confocal projection images with DAPI (blue) and CFP (cyan) channels in the contralateral SSC (i) and the thalamus (j) of animals prepared as shown in g. Scale bar, 200 μm.Data are presented as mean ± SEM. All experiments were repeated independently three times with similar results.

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