(A) A Northern blot containing RNA extracts from various adult mouse tissues (WT for Rlim) was hybridized with an Rlim probe (upper panel) and a probe recognizing β-actin as loading control (lower …
Tissue sections of mouse testes were stained using IHC with indicated antibodies. Boxed areas are shown in higher magnification. (A) DAB staining of testes sections generated from fl/Y and, as …
(A) Comparison of RLIM protein expression in mouse spleen, brain, and testis using western blot analysis. Note high levels of full-length RLIM (arrow) in testis. As loading control, the same blot …
Males systemically lacking Rlim were generated via Sox2-Cre mediated Rlim deletion (cKO/YSox2-Cre) and directly compared to fl/Y male littermates at 8 weeks of age. (A) Deletion of Rlim results in …
(A) Rlim has no essential functions during male embryogenesis and post-natal development. Using Rlimfl/KO as dames, a similar number of males out of 9 litters receiving a germline KO or the fl Rlim …
(A) Robust staining of most/all nuclei of epithelial cells lining epididymal tubules in IHC on a cross-section through the Cauda region of fl/Y males using Rlim antibodies. cKO/YSox2-Cre sections …
Cauda epididymal sperm were collected via swim-out from 8-week-old males (n = 7 fl/Y ▪; n = 9 cKO/YSox2-Cre
). (A) Sperm morphology was assessed by light microscopy. Representative images of …Sperm were incubated at 37°C for 60 min in m-TYH (non-capacitating conditions, NC) or in m-TYH supplemented with 15 mM NaHCO3 and 5 mg/ml BSA (capacitating conditions, (C)). (A) After 60 min of …
(A) Cauda epididymal sperm were collected from 8-week-old fl/Y or cKO/YSox2-Cre mice and polar metabolites were determined via LC-MS. Samples were run and data analyzed by the Metabolite Profiling …
Analyses of testicular sperm isolated from 8 weeks-old fl/Y or cKO/YSox2-Cre mice. (A) Quantification of sperm yield normalized against testis weight. n = 30, each genotype. (B) Quantification of …
(A) Examples of cKO/YSox2-Cre sperm exhibiting two axonemes (SEM). Left panel: Sperm with two axonemes. Boxed area is shown in higher magnification. Middle panel: Sperm with two axonemes and …
Animals with an Rlim cKO in the spermatogenic cell lineage or in Sertoli cells were generated via Ngn3-Cre and SF1-Cre (cKO/YNgn3-Cre and cKO/YSF1-Cre), respectively. (A) IHC on testis sections of …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Genetic reagent (M. musculus) | Rlimflox | PMID:20962847 | MGI:1342291 | |
Genetic reagent (M. musculus) | Sox2-Cre | Jackson Laboratory | JAX #008454 | PMID:14516668 |
Genetic reagent (M. musculus) | Neurog3-Cre (Ngn3-Cre) | PMID:15183725 | JAX #005667 | Dr. Andrew Leiter (UMMS) |
Genetic reagent (M. musculus) | Sf1-Cre | Jackson Laboratory | JAX #012462 | PMID:16423694 |
Antibody | Rabbit anti-Rlim (rabbit polyclonal) | PMID:11882901 | IHC (1:250) WB (1:1000) | |
Antibody | Rat anti-GATA1 (rat monoclonal) | Santa Cruz | sc265 | IHC (1:100) |
Antibody | Rabbit anti-GATA4 (rabbit polyclonal) | Abcam | ab84593 | IHC (1:500) |
Antibody | Rabbit anti-phosphoPKA (rabbit monoclonal) | Cell Signaling clone 100G7E | 9624 | WB (1:10000) |
Antibody | Mouse anti-PY (mouse monoclonal) | Millipore clone 4G10 | 05–321 | WB (1:10000) |
Antibody | Mouse anti- β-actin (mouse monoclonal) | Santa Cruz | sc47778 | WB (1:250) |
Antibody | Rabbit anti-cleaved caspase3 (rabbit monoclonal) | Cell Signaling | ab9664 | IHC (1:300) |