Journal: Aging (Albany NY)

Article Title: Age dependent increase in the levels of osteopontin inhibits skeletal muscle regeneration

doi:

Figure Lengend Snippet: ( A ) Blood sera were collected from young and old mice, either non-injured or at 3 days and 5 days after muscle injury and OPN levels were quantified by ELISA. Resting young and old blood serum levels do not show significant age-specific difference, while both at 3 days and 5 days after muscle injury old mice have higher levels of serum osteopontin than young (n=3-6, ± SEM, p**≤0.05). ( B ) Tibialis Anterior (TA) muscle sections from uninjured (resting), 3DPI and 5DPI muscle were co-immunostained for Laminin (to visualize the myofiber periphery) and OPN. OPN is undetectable in the uninjured skeletal muscle or young and old mice. In contrast, at 3 days post injury both young and old muscles show pronounced OPN at the site of injury and old muscle has considerably more OPN than young. By 5 days post injury, OPN becomes greatly diminished at the site of injury / regeneration in young muscle, but old muscle with its poor repair displays sustained OPN presence. At 3 days post injury myofibers ( C ) and myogenic stem cells ( D ) obtained from young and old mice were analyzed for OPN expression by western blotting and qRT-PCR ( F ) Old myogenic stem cells expressed higher levels of OPN as compared to young both at protein and mRNA levels, while the levels of OPN did not change with age in myofibers. (quantified in E; n=3, ± SEM, p**≤0.05).

Article Snippet: CD11b (rat monoclonal), goat normal IgG and osteopontin (goat polyclonal) for immunostaining and in vivo neutralization were from R&D, Pax7 and eMyHC was from DSHB; secondary HRP antibodies were purchased from Santa Cruz.

Techniques: Enzyme-linked Immunosorbent Assay, Muscles, Expressing, Western Blot, Quantitative RT-PCR

Journal:

Article Title: Inhibition of p53-induced apoptosis without affecting expression of p53-regulated genes

doi: 10.1073/pnas.1031695100

Figure Lengend Snippet: Verification of IL-6-induced expression of Bcl3 and Spp1. (a) Real-time quantitative PCR result of expression of Bcl3 by p53 or p53+IL-6. (b) Western blot analysis of Spp1 (osteopontin) in M1-t-p53 cells cultured at 32°C(Upper) or 37°C (Lower) without (-) or with (+) IL-6 or TG.

Article Snippet: The blots were incubated with goat anti-mouse osteopontin (Spp1) antibody (1:200) followed by horseradish peroxidase-conjugated rabbit anti-goat IgG antibody (1:2,000) (both from Santa Cruz Biotechnology).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Cell Culture

Journal:

Article Title: Inhibition of p53-induced apoptosis without affecting expression of p53-regulated genes

doi: 10.1073/pnas.1031695100

Figure Lengend Snippet: Regulation of some antiapoptotic, proapoptotic, and differentiation-associated gene expression by IL-6 and TG

Article Snippet: The blots were incubated with goat anti-mouse osteopontin (Spp1) antibody (1:200) followed by horseradish peroxidase-conjugated rabbit anti-goat IgG antibody (1:2,000) (both from Santa Cruz Biotechnology).

Techniques: Expressing

Journal: Biology of reproduction

Article Title: Osteopontin reduces polyspermy during in vitro fertilization of porcine oocytes.

doi: 10.1095/biolreprod.106.052589

Figure Lengend Snippet: FIG. 2. A) Analysis of standards of differ- ent SPP1 concentrations by Western blot- ting. B) Graphic representation of SPP1 standards. C) Analysis of SPP1 in porcine oviduct fluid. SPP1 was detected by West- ern blot analysis of the bred gilts at Day 0, 3, and 5 after estrus, and open gilts at Day 3 and 5, and depicted graphically. D) The lower molecular weight bands in the standards may represent recombinant SPP1 breakdown products.

Article Snippet: Sections were rinsed in PBS three times (10 min each) at room temperature and incubated with 10 ll goat anti-human SPP1 polyclonal antibody (1:200; R&D Systems) or normal goat serum (5 lg/ml) as control overnight at 48C.

Techniques: Western Blot, Molecular Weight, Recombinant

Journal: Biology of reproduction

Article Title: Osteopontin reduces polyspermy during in vitro fertilization of porcine oocytes.

doi: 10.1095/biolreprod.106.052589

Figure Lengend Snippet: FIG. 3. Localization of SPP1 in the porcine oviduct on Day 0 of the estrus cycle. A) Localization in the LE in the isthmus of the oviduct. B) Localization in the ampulla. (C and D), DAPI stained images. Original magnification 3400; bar ¼ 5 lm.

Article Snippet: Sections were rinsed in PBS three times (10 min each) at room temperature and incubated with 10 ll goat anti-human SPP1 polyclonal antibody (1:200; R&D Systems) or normal goat serum (5 lg/ml) as control overnight at 48C.

Techniques: Staining