Fig. 2
A non-disruptive Lrig1T2A-iCreERT2 reporter allele. a Allele design. b Comparison of recombinase variants efficiencies in a cell-based recombination assay. Mean ± standard deviation. c Targeting vector map. d Next generation sequencing sequence of the entire targeting vector. e Restriction digests of the targeting vector prepared for the electroporation. f Southern blot strategy. P, PacI. B, BamHI. K, KpnI. g Confirmation Southern blots. h SNP analysis to determine the knocked in allele. i Genotyping results of a subset of mice from heterozygote × heterozygote intercross. j Results of a Chi-square statistic calculation from intercross litters. k-k’’’’’ Summary of several behavioral parameters measured by Laboras. Asterisk, significant sample. N.s., not significant. Mean ± standard deviation. l-m’ Neurogenic markers expression in young mice. Scale bar, 10 μm. n-n” Comparison of Ascl1 and Ki-67 counts in young mice as well as Dcx+ pixel areas. Mean ± standard deviation. o-p’ Neurogenic markers expression in old mice. Scale bar, 10 μm. q-q” Comparison of Ascl1 and Ki-67 counts in old mice as well as Dcx+ pixel areas. Mean ± standard deviation. r-r’ Recombinase activity in the skin when induced with tamoxifen. s-s’ Recombinase activity in the intestine when induced with tamoxifen. A ribbon from the intestinal crypt is shown. t-t” Recombinase activity in the brain matches other Lrig1 reporter alleles. Expression is observed in the olfactory bulb, midbrain, and cerebellum. u-u’ The sfGFP-iCre-ERT2 from the allele cannot be detected by indirect immunofluorescence. Scale bar, 10 μm. u” The sfGFP-iCre-ERT2 from the allele cannot be detected by flow cytometry even though Lrig1 is expressed in the V-SVZ. v The Lrig1 protein also cannot be detected in the V-SVZ by indirect immunofluorescence. Scale bar, 10 μm. w-x However, the Lrig1 protein can be detected by flow cytometry, and is expressed in nearly all of the RFP+ cells labeled by the Lrig1T2A-iCreERT2 allele. y Numbers of singlet RFP+ cells demonstrate the kinetics of labeling with the allele. Mean ± standard deviation