TNF{alpha} mediates the skeletal effects of thyroid-stimulating hormone -- Hase et al. 103 (34): 12849 Data Supplement - HTML Page - index.htslp -- Proceedings of the National Academy of Sciences

Hase et al. 10.1073/pnas.0600427103.

Supporting Information

Files in this Data Supplement:

Supporting Figure 8
Supporting Text
Supporting Figure 9
Supporting Figure 10

Supporting Figure 8 Fig. 8.
Rescue experiments showing osteoclast formation (cells/well) (Left) and tartrate-resistant acid phosphatase (TRAP) mRNA expression (Right) in bone marrow cell cultures isolated from various genotypes, namely TSHR^+/+/TNF^+/+ (w/w), TSHR^+/-/TNF^+/+ (h/w), and TSHR^+/-/TNF^+/- (h/h) mice. *, P<0.05, comparisons with wild type (w/w) mice.

Supporting Figure 9 Fig. 9.
RAW-C3 cells were incubated with murine IL-1 (10 ng/ml), murine TNFa (50 ng/ml) alone, or in combination (IL-1/TNFa) for 30, 60, and 120 min. TNFa mRNA expression was determined by real-time PCR. *, P<0.05, comparisons with vehicle (Cont).

Supporting Figure 10 Fig. 10.
Cyclic AMP (cAMP) levels in cell lysates from empty-C3, TSHR-C3, or caTSHR-C3 cells or CHO-TSHR, which were stimulated with thyroid-stimulating hormone (TSH) (10 milliunits per ml) for 2 h or forskolin (10 m M) for 30 min. Data show means of 2–4 samples.

Supporting Text cAMP Measurement.
cAMP levels were determined in TSHR-C3 and caTSHR-C3 cells and CHO cells overexpressing hTSHR-EGFP (CHO-TSHR) (provided by Ratif Rauf, Mount Sinai School of Medicine, New York, NY). Cells were stimulated with bovine TSH (10 milliunits per ml) for 2 h or forskolin (10 mM) for 30 min, and cAMP levels in the cell lysates were determined by a cAMP Enzymeimmunoassay Biotrack (EIA) kit (Amersham Pharmacia, Piscataway, NJ).