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. 2021 Mar 29;10:e64393. doi: 10.7554/eLife.64393

Figure 2. Sclerostin protein is rapidly degraded after PTH exposure in vitro and ex vivo.

(A) Ocy454 cells transfected with GFP-sclerostin were treated with vehicle or PTH (1–34) (10 nM) for the indicated time and were lysed. Western blots were probed for sclerostin and β-actin (n = 2–3). (B) Dissected tibiae flushed of marrow were treated with vehicle or PTH (1–34) (10 nM) for 30 min ex vivo and homogenized. Western blots were probed for sclerostin and β-actin (n = 6 mice). (C) Ocy454 cells were treated with vehicle or PTH (1–34) (10 nM) for the indicated time and were lysed. Western blots were probed for pCaMKII and total CaMKII (n = 6–8). Graphs depict mean ± SD. *p<0.05, **p<0.01 by two-way ANOVA with Holm–Sidak post hoc correction (A, C) or unpaired two-tailed t-test (B).

Figure 2.

Figure 2—figure supplement 1. Acute treatment with PTH does not affect Sost mRNA hours later.

Figure 2—figure supplement 1.

Ocy454 cells were treated with PTH (10 nM) for 30 min and then collected 5.5 hr after treatment to examine Sost mRNA levels as normalized to Gapdh, Rpl13, and Hprt by RT-qPCR (n=6).