Extended Data Fig. 6: Validating the role of Cplx1 in ultrasound perception of mouse ACx.
From: Complexin-1 enhances ultrasound neurotransmission in the mammalian auditory pathway
(a) Fluorescence-labeled siRNA transfection in P19 cell line. Representative images from 3 independent experiments. (b) Relative normalized expression of Cplx1 in 3 different Cplx1 siRNA transfected P19 cell line detected by RT-qPCR. n = 3 biologically independent samples. (c) Example micrographs showing the range of AAV infection labeled by eGFP. Green regions (middle) represent the ACx. Representative images from 4 independent experiments. (d) RT-qPCR validation of KD efficiency of Cplx1 in mouse ACx. Two (Cplx1-KD) or three (NC) biological replicates, each with three technical repeats, were conducted in each sample. Data are mean values ± SD. Two-sided t-test P values are indicated. (e) Representative example of locomotion of NC and Cplx1 KD mice tested with 16 kHz or 63 kHz cue in context B box. Dots indicate the ___location of the mouse every 0.04 s. (f) Relative normalized expression of Cplx1 and smCplx1 in rescued mice by bulk RNA-seq analysis. n = 4 biologically independent samples. (g) UMAP visualization of mouse ACx inhibitory neurons based on a previous study and the expression patterns of Pvalb and Cplx1. (h) Representative flow cytometry results of sorting mCherry+ cells in PV-ires-Cre mouse ACx. (i) RT-qPCR validation of specific KD efficiency of Cplx1 in PV-ires-Cre mouse ACx. Two biological replicates, each with three technical repeats, were conducted in each group. Data are mean values ± SD. Two-sided t-test P values are indicated. (j) Fluorescence-labeled sgRNA transfection in P19 cell line. Representative images from 3 independent experiments. (k) T7EI assays for assessing the CRISPR/Cas9 mediated indels formation of Cplx1 in P19 cell line. Representative images from 2 independent experiments.
