Supplementary Figure 1: Experimental timeline and post-experiment histology.

(a) Experimental timeline. (b) Post-study representative confocal photomicrograph examples from two individual PV-Cre mice used in this study. Each example includes merged images along with separate channel images for GCaMP6f (green), tdTomato (red), and anti-PV immunofluorescence (blue). White arrows indicate cells with co-localized expression of tdTomato and anti-PV. Red arrows highlight cells expressing anti-PV that were not tdTomato labeled. (c) Box plots showing quantification of viral specificity from a subset of PV-Cre mice recorded from cohort 1 and cohort 2 of this study (n = 7; see methods). 74.0 ± 0.8% (mean ± s.e.m.) of PV immunoreactive cells co-expressed tdTomato and 96.1 ± 0.2% (mean ± s.e.m.) of tdTomato expressing cells were co-immunoreactive for PV. (d) Same as (b), but for two representative Chat-Cre mice, with anti-Chat immunofluorescence in blue. (e) Box plots showing quantification of viral specificity from a subset of Chat-Cre mice recorded from cohort 1 and cohort 2 of this study (n = 7; see methods). 78.8 ± 0.4% (mean ± s.e.m.) of Chat immunoreactive cells co-expressed tdTomato and 97.2 ± 0.2% (mean ± s.e.m.) of tdTomato expressing cells were immunoreactive for Chat. In a separate analysis, comparison of tdTomato images taken during all imaging sessions (n = 28 sessions in 6 PV-cre mice, and 6 Chat-cre mice), revealed that 63.2 ± 5.1% (mean ± s.e.m.) of all tdTomato positive cells, also co-expressed GCaMP6f. For all box plot figures, middle lines indicate the median, lower and upper edges of the box indicate quartiles below and above the median, and upper and lower whiskers indicate maximum or minimum values respectively.