Figure 7.

Silencing of NOX3 in vivo. NOX3 siRNA #248 was manually delivered to the inner ear through the posterior semicircular canal or into the middle ear through the tympanic bulla. (A) Bar graph showing the mRNA level (qPCR) of Nox3 and p22^phox (Cyba) 3 days following inner ear delivery (canalostomy) (n = 6). (B) Bar graph showing the mRNA level (qPCR) of Nox3 and p22^phox (Cyba) 3 days following middle ear delivery (n = 2). (A,B) mRNA level in the siRNA delivered cochlea was normalized to the contralateral cochlea (100%). The data represent the average +- SEM. (C) Representative images corresponding to the Rosenthal's canal in situ hybridization (BROWN assay) and hematoxylin counterstaining (C1). Dapb (dihydrodipicolinate reductase) probe was employed as negative control (–), while Ppib (Peptidyl-Prolyl Cis-Trans Isomerase was employed as positive control (+). DAB channel, corresponding to the mRNA signal, was processed in a fluorescent like manner (LUT applied), in order to facilitate visualization (C2). (D) Bar graph representing RNAscope (in situ hybridization) Nox3 mRNA signal quantification in the Rosenthal's canal area. Nox3 signal was normalized to the Rosenthal's area (mm²). The data represents the average +- SEM of Nox3 signal quantified from five to eight cochlear slices / cochlea from three animals.