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. 2022 Apr 4;10:816764. doi: 10.3389/fcell.2022.816764

FIGURE 4.

FIGURE 4

SSHTN mice showed increased TNF-α and activation of bone RAS in vivo. (A) IL-1β mRNA expression levels in normal control and SSHTN mice. (B) TNF-α mRNA expression levels in normal control and SSHTN mice. (C) AGTR1 mRNA expression levels in normal control and SSHTN mice. (D) ACE mRNA expression levels in normal control and SSHTN mice. IL-1β, TNF-α, AGTR1, and ACE mRNA levels were measured by real-time RT-PCR. Total RNA was isolated from the tibiae of normal control and SSHTN mice. Expression levels of IL-1β, TNF-α, AGTR1, and ACE mRNA were normalized relative to GAPDH (n = 4). (E) Western blot analysis bands showing the expressions of TNF-α and AGTR1 protein in the distal femur of normal control and SSHTN mice. (F) Relative protein expression of TNF-α and AGTR1 were measured using ImageJ software (n = 4). β-Actin was used as a loading control. (G) Serum levels of TNF-α was determined using ELISA MAX Standard Set Mouse TNF-α. Serum samples were collected from normal control and SSHTN mice (n = 4).