Journal: Cellular and molecular biology (Noisy-le-Grand, France)
Article Title: Pro-inflammatory GPR75 and anti-apoptotic phospholipase signaling pathways contribute to the ameliorating effect of soluble epoxide hydrolase inhibition on chronic experimental autoimmune encephalomyelitis in mice.
doi: 10.14715/cmb/2023.69.10.2
Figure Lengend Snippet: Figure 3. TPPU increased the PLC β4, PI3K p85α, Akt1, MEK1/2, ERK1/2, CREB1, and Bcl-2 expression and/or activity induced by MOG35-55 peptide/PT in the (a, b, c, d, e, f, and g) brain and (h, i, j, k, l, m, and n) spinal cord of PBS-, DMSO-, or TPPU-injected mice. The expression of unphosphorylated and phosphorylated pro teins for PLC β4, PI3K p85α, Akt1, MEK1/2, ERK1/2, CREB1, and Bcl-2 in tissues was measured by immunoblotting technique. Data are expressed as means ± SEM of 3-4 animals. + P<0.05 versus PBS- injected mice (EAE control group); # P<0.05 versus DMSO-injected mice (vehicle control group).
Article Snippet: Nitrocellulose membranes containing transferred proteins from the gel were blocked with nonfat dry milk in Tris-buffered saline and then incubated with specific antibodies against (1) sEH (13560; Cayman), (2) GPR75 (GTX55193; GeneTex, Hsinchu City, Taiwan), (3) Gαq/11 (sc-515689; Santa Cruz Biotechnology, Santa Cruz, CA, USA), (4) c-Jun (sc-1694; Santa Cruz), (5) phosphorylated c-Jun (p-c-Jun) (sc-16312; Santa Cruz), (6) PLC β4 (sc-16631; Santa Cruz); (7) PI3K p85α (OAAF05915; Aviva Systems Biology, San Diego, CA, USA), (8) phosphorylated PI3K p85α (p-PI3K p85α) (OAEC00301; Aviva Systems Biology), (9) Akt1 (sc-271149; Santa Cruz), (10) phosphorylated Akt1 (p-Akt1) (sc-52940; Santa Cruz), (11) MEK1/2 (sc-81504; Santa Cruz), (12) phosphorylated MEK1/2 (p-MEK1/2) (sc-81503; Santa Cruz), (13) ERK1/2 (sc-514302; Santa Cruz), (14) phosphorylated ERK1/2 (p-ERK1/2) (sc-136521; Santa Cruz), (15) CREB1 (sc-271; Santa Cruz), (16) phosphorylated CREB1 (p-CREB1) (sc-81486; Santa Cruz), (17) Bcl-2 (sc-7382; Santa Cruz), (18) myelin PLP (bs-11093R-HRP; Bioss Antibodies Inc., Woburn, MA, USA), and (19) SEMA3A (sc-74554; Santa Cruz) in bovine serum albumin overnight at 4°C.
Techniques: Expressing, Activity Assay, Injection, Western Blot, Control