Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - Two transcripts for rat CRISP4 detected in all regions of the epididymis. Northern blot analysis probing for rat Crisp4 (A) and GAPDH loading control (B) expression in the caput, corpus, and cauda epididymis. A major (~1.4 kb) and a minor (~950 bp) transcript were detected for rat CRISP4 in each region of the epididymis. (C) Densitometric analysis of band intensity normalized to GAPDH intensity demonstrates that the 1.4 kb band is ~4X more abundant than the 950 bp band. Each lane represents 300 ng Poly A+ RNA (Oligotex mRNA isolation kit, Qiagen, Valencia, CA). Probe templates were a 600 bp EcoRI fragment of the rat Crisp4 cDNA and the 905 bp DECAtemplate-GAPDH-mouse (Ambion, Austin, TX). Relative RNA size markers are indicated to the left of each panel (Promega, Madison, WI).
• Supplemental Figure 2 - Rat CRISP1, but not CRISP4, is sensitive to PNGase F-mediated deglycosylation. The “no-detergent soluble” protein extracts from the cauda epididymis shown in Figure 4 were subjected to PNGase F-mediated removal of N-linked glycosylations according to the manufacturer’s instructions (N-Glycanase PLUS, Prozyme, San Leandro, CA). Western blotting of the samples probing for either CRISP1 (CAP-A antibody) (A) or CRISP4 (B) reveals that N-glycanase treatment shifts the apparent MWR of CRISP1 but does not alter the MWR of rat CRISP4 in the cauda epididymis.