Journal: The Journal of Biological Chemistry

Article Title: Retinal Degeneration Slow (RDS) Glycosylation Plays a Role in Cone Function and in the Regulation of RDS·ROM-1 Protein Complex Formation *

doi: 10.1074/jbc.M115.683698

Figure Lengend Snippet: Knockin of the N229S mutation removes RDS glycosylation while preserving normal RDS message and trafficking. A, the N229S mutation was introduced to exon 2 of the Rds gene to drive expression of unglycosylated RDS in the native regulatory environment. B, at P30, total RNA was isolated from murine retinas of the indicated genotypes, analyzed by quantitative RT-PCR for RDS, and normalized to the housekeeping gene HPRT. N.S., not significant. C, Northern blotting with radiolabeled random primed RDS cDNA as a probe was performed on equivalent RNAs in B. 28S and 18S ribosomal RNAs were used as a size reference, and 28S is shown in the bottom panel. D, retinal extracts from WT and rdsN/N were subjected to enzymatic deglycosylation by PNGase F. Actin reactivity was used as a loading control. IB, immunoblot. E, retinal cross-sections at P30 from WT and rdsN/N were immunolabeled with antibodies for S-opsin (green), Na+K+ATPase (blue), and either RDS (red, left), ROM-1 (red, center), or rhodopsin (red, right). Nuclei were counterstained with DAPI (gray). IS, inner segment; OPL, outer plexiform layer. Scale bar = 20 μm.

Article Snippet: Antibodies The antibodies that were used in this study were as follows: RDS-CT (rabbit polyclonal, generated in-house, 1:1000) for immunofluorescence (IF) ( 18 , 31 ), RDS-2B7 (mouse monoclonal, generated by Precision Antibodies, 1:1000) for Western blot (WB) ( 31 , 38 ), ROM1-CT (rabbit polyclonal, generated in-house, 1:1000) for WB ( 14 ), ROM1–2H5 (mouse monoclonal, generated in-house,1:10) for WB ( 32 ), Na + K + ATPase (mouse monoclonal, Developmental Studies Hybridoma Bank, developed under the auspices of the NICHD/National Institutes of Health and maintained by the University of Iowa Department of Biology (Iowa City, IA), 1:500) for IF, Rhodopsin-1D4 (mouse monoclonal, provided by Dr. Robert Molday, University of British Columbia, 1:1000) for IF and WB, S-opsin (goat polyclonal, N-20, Santa Cruz Biotechnology (Santa Cruz, CA), 1:500) for IF, and β-actin (HRP-conjugated mAb, Sigma-Aldrich (St. Louis, MO) 1:10,000) for WB.

Techniques: Knock-In, Mutagenesis, Glycoproteomics, Preserving, Expressing, Isolation, Quantitative RT-PCR, Northern Blot, Random Primed, Control, Western Blot, Immunolabeling

Journal: Materials

Article Title: Stochastic Finite Element Analysis Framework for Modelling Mechanical Properties of Particulate Modified Polymer Composites

doi: 10.3390/ma12172777

Figure Lengend Snippet: Schematic illustrating the Random Number Generator (RNG) module .

Article Snippet: The Random Number Generator module (RNG, see ) was developed using the general mathematical programming environment MATLAB, enabling the generation of random numbers required for input variables.

Techniques: