Fig 2: Activation of P2Y6R by UDP is partially dependent on ERK1/2 phosphorylation. a Representative western blots showing that ERK1/2 phosphorylation was stimulated in a time-dependent manner by UDP. b Representative western blots and RT-PCR showing that COX-2 protein and MIP-2 mRNA levels were decreased in UDP-stimulated BV-2 cells pretreated with U0126 (ERK1/2 inhibitor). c ELISA showing that MIP-2 secretion level was decreased in UDP-stimulated BV-2 cells pretreated with U0126. SB (SB203580, P38 inhibitor), SP (SP600125, JNK1/2 inhibitor). Data are shown as the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 vs Con, ## p < 0.01, ### p < 0.001 vs LPS

Fig 3: High fat diet activated Ang II-AT1R axis in rats.A. Protein expressions of kinases downstream of the Ang II-AT1R axis, p38 and ERK. B. Gene expression of Nox4 in adipose tissue. *p < 0.05 vs WT-ND rats. Data were presented as Mean ± S.E.M. n = 4.

Fig 4: PRMT1 acts downstream of M KK3 (MAPK kinase 3) to promote erythroid differentiation, and the R49/149K non-methylation mutant exhibits a reduced interaction with MKK3. MKK3 and MKK6 were knocked down (A) and (B) as described in the Methods section. AraC-induced erythroid differentiation was significantly suppressed in MKK3-knockdown (KD1 and KD2) cells (A); however it was stimulated in MKK6-knockdown (KD1 and KD2) cells (B). The activation of p38 was significantly reduced in MKK3 KD1 and KD2 cells (C). Ectopic expression of p38a, but not p38ß, in MKK3 KD1 cells could partially rescue differentiation (D). However, the R49K, R149K and R49/149K mutants lost the ability to promote differentiation (E). Ectopic expression of HA-PRMT1 promoted differentiation in parental K562, Luc and MKK3 KD1 and KD2 cells but had no effect in MKK6 KD1 and KD2 cells (F). Luc is the vector control cell. To examine the interaction of p38a with MKK3 and MKK6, Flag-p38a wild type and R49/149K mutant proteins were expressed in p38a KD cells. After cells were stimulated with AraC (1 µM) for 5 h, Flag-p38a proteins were immunoprecipitated and the protein levels of MKK3 (G) or MKK6 (H) in the immunoprecipitates were examined by Western Blot. The levels of MKK3, pp38 and p38 were quantified by Multi-Gauge V3.0 analysis. All results shown are representatives of three independent experiments. Statistical analysis was performed with results from three separate experiments. *** p < 0.005. ** p < 0.01.

Fig 5: Proposed differential signaling schema comparing PNFs and CNFs. Although both RAS/MEK/ERK and RAS/MKK3/P38 signaling occurs in both tumor types, RAS activation results in differential signaling strength and fidelity through RAS/MEK in PNFs (left panel) and, alternatively, RAS/MKK3/P38 in CNFs (right panel). CNFs divert RAS activation through MKK3 thereby enhancing the cellular response to stress and inflammation that is mediated by MKK3/p38, whereas the downstream impact in RAS/MEK dependent PNFs is unchecked growth and proliferation (left). CNFs still maintain growth and proliferation signaling through RAS/ERK or p38/ERK, but strongly exhibit a pro-inflammatory phenotype and are characterized by broad chromatin remodeling (right)