hRHO-C110R

Fig.1 Detection of RHO expression in testis by RT-PCR. 

Wild type: one band at 351 bp with primers F1/R1(mRho) and one band at 123 bp with primers F/R(mGapdh).

Homozygous: one band at 492 bp with primers F2/R2(hRHO) and one band at 123 bp with primers F/R(mGapdh).

Abbr. M, DNA marker; HO, homozygous; WT, wild type.

Fig.2 RNA sequencing analysis of RT-PCR products.

Abbr. HO, homozygous.

Fig.3 Representative images of retinal morphology in WT C57BL/6 mice, hRHO mice, and homozygous hRHO-C110R mice. The results showed that the homozygous hRHO mice (n=2, 14 weeks, male) had no defect in morphology of fundus and retinal layers compared to the WT C57BL/6 mice. Compared with WT C57BL/6 and hRHO mice, the outer nuclear layer (ONL) of hRHO-C110R (n=2, 23 weeks, male) mice was thinner. 

Fig.4 Representative images of HE staining in WT C57BL/6 mice, hRHO mice, and homozygous hRHO-C110R mice. The results indicated that the photoreceptor outer segments (OS), outer nuclear layers (ONL), and inner nuclear layer (INL) became thinner in hRHO-C110R mice (n=2, 23 weeks, male) compared with WT C57BL/6 and hRHO mice (n=2, 14 weeks, male). Scale bar, 50 μm.

Abbr：OS, outer segment; ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer. 

Fig.5 Representative images of immunohistochemistry staining in WT C57BL/6 mice, hRHO mice, and homozygous hRHO-C110R mice. The results indicated that the outer nuclear layers (ONL) of homozygous hRHO-C110R mice (n=2, 23 weeks, male) was thinner, and the expression of rhodopsin protein on the retina was abnormal compared with WT C57BL/6 and hRHO mice (n=2, 14 weeks, male). Black arrow represents the positive area of rhodopsin protein. Scale bar, 50 μm.

Abbr：OS, outer segment; ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer. 

Fig.6 Electroretinogram (ERG) responses in WT C57BL/6 mice, hRHO mice, and homozygous hRHO-C110R mice. ERG measurements showed that the amplitudes scotopic a-wave (A), scotopic b-wave (B), photopic a-wave(c), and photopic b-wave (D) significantly decreased in hRHO-C110R (n=2, 23 weeks, male) mice compared with WT C57BL/6 and hRHO mice (n=2, 14 weeks, male).