Figure 2

Relative expression of GFP in Rosa26^tm(CAG-LSL-Tmc2-IRES-GFP) mice. Total RNA was prepared from single cochlea harvested from Rosa26^tm(CAG-LSL-Tmc2-IRES-GFP)];Tmc1^Δ/ΔTmc2^Δ/+ positive for Gfi1^cre (Tm[Tmc2]) or negative for Gfi1^cre (Cre-neg) at P1, P8 and P20. Quantitative RT-PCR was performed with β-actin and GFP primers in triplicate (see methods) and repeated from biological replicates (number of samples are noted above the box plots). Due to bicistronic expression of Tm[Tmc2] and GFP in Rosa26^tm(CAG-LSL-Tmc2-IRES-GFP) mice, the expression level of GFP reflected Tm[Tmc2] expression. The cycle threshold (Ct) for each sample was normalized relative to the Ct value of β actin from the same sample. Relative expression was calculated by normalization to P1 Gfi1^Cre negative control samples. Significant expression of the knockin gene was detected at each stages from P1 to P20. Data are represented in box plots as means ± S.E.M. Statistical analysis were performed using the independent t-test. *p < 0.05; **p < 0.01; ***p < 0.001.