Journal: Scientific Reports

Article Title: Proteomics identifies multiple retinitis pigmentosa associated proteins involved in retinal degeneration in a mouse model bearing a Pde6b mutation

doi: 10.1038/s41598-024-72821-1

Figure Lengend Snippet: Western blots validation of RP related DEPs in rd10 mice ( n = 3 retinas per group). (A) (E) (F) Proteins levels of Pde6a and Prph2 in age-matched WT and rd10 retinas. (B) (G) (H) (I) Bbs2, Bbs7 and Nr2e3 expression in retinas at 5 or 8 weeks. (C) (J) (K) Proteins expression of Rho and Rom1 in WT and rd10 mice retinas. (D) (L) (M) (N) (O) Rbp3, Tulp1, Bbs4 and Arl6 protein levels in age-matched mice retinas. 3 repeated lanes were conducted for each group. The results were mean ± SEM. significance of difference(** p < 0.01, *** p < 0.001, **** p < 0.0001) was determined by using unpaired t-test.

Article Snippet: A 1.5 h soak in 5% non-fat milk blocked PVDF membranes was followed by an overnight incubation with antibodies against Pde6b (PA1-722, Thermo Fisher Scientific), Pde6a (PA1-720, Thermo Fisher Scientific), Tulp1 (18971-1-AP, Proteintech, Chicago, USA), Prph2 (18109-1-AP, Proteintech), Rbp3 (14352-1-AP, Proteintech), Bbs2 (11188-2-AP, Proteintech), Bbs7 (18961-1-AP, Proteintech), Bbs4 (ER1904-15, Huabio), Nr2e3 (14246-1-AP, Proteintech), Rho (ET1704-12, Huabio), Rom1 (CSB-PA344843ESR2HU, Cusabio, Wuhan, China), Arl6 (12676-1-AP, Proteintech) and β-actin (ab8227, Abcam) at 4 °C. β-actin was used as the endogenous reference.

Techniques: Western Blot, Biomarker Discovery, Expressing

Journal: bioRxiv

Article Title: A spontaneous mutation in ADIPOR1 causes retinal degeneration in mice

doi: 10.1101/2024.06.06.597783

Figure Lengend Snippet: Abnormal levels of Rbp3 mRNA and altered spatial distribution of RBP3 protein. ( A ) Rbp3 mRNA is significantly upregulated in postnatal day (PD) 15 AdipoR1 −/− mice (KO) by RNAseq compared to AdipoR1 +/− (HET) and AdipoR1 +/+ (WT). p < 0.0001. Another interphotoreceptor matrix gene, Impg2 , is similar among the genotypes. p = 0.4614. ( C ) RBP3 (aka IRBP) protein in PD30 AdipoR1 +/+ is observed in two bands, one in the inner segments (IS) and the other at the tip of the OS, bordered by the outer nuclear layer (ONL) and (RPE). ( D ) In contrast, RBP3 is broadly distributed in the is and os region in Adipor1 −/− mice ( n = 3 per genotype). Scale bar is 10 μm for C and D.

Article Snippet: The primary antibody for RBP3 (also known as IRBP) was rabbit anti-RBP3 polyclonal (Proteintech, Catalog No. 14352-1-AP, Illinois).

Techniques: