Supplementary Figure 9: Optogenetic activation of VIP BLA interneurons does not induce associative fear learning.

a, Schematic illustrating the entire behavioral paradigm used for optogenetic gain-of-function experiments, including details about CS, US and blue light pattern applied. b-c, Representative example images of bilateral expression of b, ChR2-EYFP and c, GFP in VIP interneurons in the BLA of VIP-cre mice with corresponding optical fiber placement (dashed lines). Scale bar, 200 µm. d, Position of optical fiber tips (squares) and maximum virus spread (shades) in all mice included in optogenetic experiments matched to a mouse brain atlas. LA, lateral amygdala; BA, basal amygdala; CEA, central amygdala. e, CS presentations on habituation day do not induce freezing in naïve mice. f-g, Optogenetic activation of VIP BLA interneurons has no effect on f, freezing or g, running speed during or after blue light stimulus presentation in naïve mice (ISI, inter-stimulus interval). h, Optogenetic activation of VIP BLA coupled to CS presentations does not induce associative fear learning as measured by freezing responses to the CS on retrieval day. i, Similarly, no difference is detected in the average speed during the CS between groups. j, Optogenetic activation of VIP BLA interneurons for 2 s or 10 s on a separate laser test day does not induce freezing behavior. For panels d-j: ChR2, N=8 mice; GFP, N=9. Confocal images are representative for N=8 mice (b), N=9 (c). Box-and-whisker plots show median values and 25^th/75^th percentiles with 10^th to 90^th percentile whiskers, dots additionally indicate the mean. Details of statistical analysis are listed in Supplementary Table 3.