Fig 2: Schematic diagram of LRRC4 as a tumor suppressor in glioblastoma. The MEK/ERK/MAPK pathway is one of the most frequently aberrantly activated signaling pathways, and LRRC4 is an important tumor suppressor and has decreased expression in glioblastoma. Ectopic expression of LRRC4 abrogates the MEK1/2-ERK1/2 interaction. LRRC4 competitively inhibits the binding of ERK1/2 with MEK1/2 and prevents the phosphorylation of ERK1/2 and nucleus translocation. This further suppresses the ERK-mediated activation of the downstream transcripts to inhibit cell proliferation and invasion. In glioblastoma, the decrease or loss of LRRC4 failed to block the ERK 1/2-MEK 1/2 interaction and prevents MEK1/2 activation of ERK1/2

Fig 3: ADAM9 affects auto-/paracrine MEK/ERK signaling. (A) Western blot of phosphorylation of MEK1/2 (S217/221) and ERK1/2 (T202/204) in AsPC-1 and MiaPaCa-2 cells following ADAM9 expression silencing. In AsPC-1 cells, we observed decreased pFAK (Y397), which was absent in MiaPaCa-2 cells. Quantitation of pFAK, pMEK1/2, and pERK1/2 in AsPc-1 and MiaPaCa-2 cells from three independent experiments. (B) Western blot analysis shows decreased phosphorylation of MEK1/2 (S217/221) and ERK1/2 (T202/204) in HUVEC cells following 30 min of incubation with cancer cell conditioned medium from both AsPC-1 and MiaPaCa-2 cells.

Fig 4: Decreased metabolic and growth kinases in crSTIM1-/- hearts. Immunoblots and densitometric analysis of AMPK (a), PDPK1 (b), ERK1/2 (c), and MEK1/2 (d), phosphorylation in hearts isolated from control and crSTIM1-/- mice treated subcutaneously with 15 min saline or phenylephrine. Protein expression is normalized to total protein and/or GAPDH. STIM1, stromal interaction molecule 1. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, versus age-matched control (ANOVA/ Tukey's post hoc), n = 3 per genotype

Fig 5: Role and mechanism of MFAP4 in cardiac fibroblasts A, Polymerase chain reaction analysis of mRNA level of MFAP4 (microfibrillar-associated protein 4) between cardiomyocytes and cardiac fibroblasts (n=6). The data were compared by unpaired Student t-test. B and C, Polymerase chain reaction analysis of the mRNA expression level of MFAP4 and markers of, hypertrophic and fibrosis markers when in cardiomyocytes and cardiac fibroblasts were stimulated by angiotensin II (Ang II), phenylephrine, and or TGF-ß, respectively (n=6). (D through G) PCR analysis of the mRNA expression levels of aSMA, Col1a, Col3, and fibronectin (n=6). H, Immunofluorescence staining of aSMA (n=6). I through O, Representative blots and quantitative results for P-FAK/T-FAK, P-PI3K/T-PI3K, P-AKT/T-AKT, P-MEK1/2/T-MEK1/2, P-ERK1/2/T-ERK1/2 protein expression in cardiac fibroblasts each group (n=4). P through T, Polymerase chain reaction analysis of the mRNA expression levels of MFAP4 and fibrosis markers in cardiac fibroblasts treated with Ad-MFAP4 and an inhibitor of FAK (PND-1186) (n=6). Ang II indicates angiotensin II; CF, cardiac fibroblasts; CMs, Cardiomyocyte; Col1a, collagen Ia; Col3, collagen 3; CTGF, connective tissue growth factor; ERK, extracellular signal regulated kinase; FAK, focal adhesion kinase; ; MEK, mitogen-activated protein kinase (MAPK)/ERK kinase; MFAP4, microfibrillar-associated protein 4; PI3K, phosphatidylinositol 3kinase; PSR, Picrosirius red; TGF-ß, transforming growth factor-ß; and aSMA, alpha-smooth muscle actin. *P<0.05; **P<0.01; *P<0.001.