Detection of native PfN23.
A, tissue-specific gene expression of PfN23 by RT-PCR analysis. Total RNA extracted from mantle edge (ME), mantle pallial (MP), gonad (Gon), foot, viscus (Vis), hemocytes (Hem), and adductor muscle (Mus) was used for RT-PCR. RT-PCR without a template was used as a negative control (Control). The housekeeping gene GAPDH was used as a positive control. B–E, detection of native PfN23 in shells by immunogold staining of the nacreous layer surface. B, SEM image of immunogold staining of the nacreous layer surface. D, SEM image of the control nacreous layer surface. C and E, back-scattered electron mode scanning electron microscope (SE-BSE) image of the same area shown in (B and D), respectively. Scale bars, 5 μm in B–E.