Fig. 5.

CTMP induced by DIO inhibits Akt phosphorylation and attenuates innate immune responses. (A) Total protein was extracted from un-stimulated BMMΦ isolated from obese (OB) and lean (LN) mice. CTMP, PP2A, and PTEN were detected by using Western blot. (B) CTMP suppresses Akt phosphorylation. THP-1 cells were stably transfected with HA-tagged CTMP or empty vector and then infected with P. gingivalis for 0, 4, and 24 h, and Western blots were performed using total cell lysates. (C–E) CTMP inhibits innate immune responses. THP-1 cells stably transfected with CTMP or empty vector were infected with P. gingivalis for 0, 4, and 24 h; the induction of iNOS was detected in cell lysates by Western blot (C), and the release of TNF-α (D) or IL-10 (E) was measured by ELISA using cell culture supernatants. (F and G) CTMP physically interacts with Akt. THP-1 cells were stably transfected with CTMP or empty vector. After serum starvation (24 h), total proteins were extracted and immunoprecipitated using either anti-HA (F) or anti-CTMP (G) antibodies; Akt was analyzed by using an antibody against total Akt by Western blot. Each image represents one of the triplicate results. Data are expressed as mean ± SEM (n = 3; *P < 0.05).