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. 2024 Apr 8;65(4):18. doi: 10.1167/iovs.65.4.18

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Copyright 2024 The Authors

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Figure 1. — Generation of transgenic mice and verification of the Slc4a11 knockdown. (A) Schematic of the gene animals. The tamoxifen-inducible knockdown of Slc4a11 mouse (B6.Slc4a11^Flox/Flox/Rosa^Cre-ERT2^/^Cre-ERT2) was crossed either with Col8a2 wildtype (B6.Col8a2^+/+) or Col8a2 (Q455K) knock-in (B6.Col8a2^ki/ki) animals to produce litters with the genotype B6.Slc4a11^Flox/Flox/Rosa^Cre-ERT2^/^Cre-ERT2/Col8a2^+/+ or B6.Slc4a11^Flox/Flox/Rosa^Cre-ERT2^/^Cre-ERT2/Col8a2^ki/ki referred to as single mutants (SMs) and double mutants (DMs), respectively. The animals were either fed with tamoxifen (Tm) for 2 weeks at 5 weeks of age - SM (+Tm) or DM (+Tm), or fed with standard chow - SM (−Tm) or DM (−Tm). Age-matched wildtype animals were also fed with Tm for 2 weeks. (B) Quantification of Slc4a11 transcripts using kidney lysates from experimental SM (+Tm) or DM (+Tm), control SM (−Tm) or DM (−Tm), and wildtype (WT) animals at 16 weeks of age (n = 3 to 4 replicates). (C) The cDNA was used to perform PCR to confirm the knockdown of Slc4a11 (floxed band = 438 bp and wildtype band = 353 bp) with GAPDH as a loading control. Mean ± standard deviation (SD), ns = not significant, *P < 0.05, **P < 0.01, ****P < 0.0001 (1-way ANOVA with Uncorrected Fisher's LSD multiple comparisons). B6.Slc4a11^Flox/Flox/Rosa^Cre-ERT2^/^Cre-ERT2/Col8a2^+/+ fed with Tamoxifen – SM (+Tm), B6.Slc4a11^Flox/Flox/Rosa^Cre-ERT2^/^Cre-ERT2/Col8a2^+/+ fed with normal chow – SM (−Tm), B6.Slc4a11^Flox/Flox/Rosa^Cre-ERT2^/^Cre-ERT2/Col8a2^ki/ki fed with Tamoxifen – DM (+Tm), B6.Slc4a11^Flox/Flox/Rosa^Cre-ERT2^/^Cre-ERT2/Col8a2^ki/ki fed with normal chow – DM (−Tm).