Fig 2: EGFR and CCNE1 amplifications could be potential biomarkers of therapy targeting EGFR and CDK2. a The efficacy of BK011 and cetuximab on five PDX models (n = 5 per group). Tumor volumes and proportion of tumor growth inhibition were expressed as means ± SD. NS, p > 0.05; ***p < 0.001 according to repeated measures ANOVA. CR, complete regression. b The immunohistochemical analysis of EGFR and phosphorylated EGFR expression in five PDX models. Scale bar represents 100 μm. c The expression and phosphorylation of EGFR, AKT, and S6 after BK011 or cetuximab treatment assessed by immunoblot. d CCNE1 copy numbers of eight PDX models quantified by real-time PCR and the tumor growth inhibition of AZD5438 in four PDX models. The copy number and proportion of tumor growth inhibition were expressed as means ± SD. NS, p > 0.05; ***p < 0.001 according to repeated measures ANOVA. e AZD5438 showed potent antiproliferative activity in two PDX models with CCNE1 copy numbers ≥ 29 alterations (n = 5 per group). Tumor volumes were expressed as means ± SD. NS, p > 0.05; ***p < 0.001 according to repeated measures ANOVA. f AZD5438 exerted antitumor effect in case 168 accompanied by reduction of expression of CDK2, CCNE1, and phosphorylated retinoblastoma (pRb). Whereas in case number 111, slight reduction of CDK2 but no impact on level of CCNE1 or pRB

Fig 3: State of phosphorylation of protein kinases. (A) Representative images of phospho-kinase array blots assayed with protein extracts from MCF-7 cells, indicative the state of phosphorylation of multiple kinases was assayed using the Proteome ProfilerTM Array and protein extracts fromMCF-7 cells treated for 6 h with doxorubicin (1 μM) and/or melatonin (1 nM). The white oval remarks the dots corresponding to p70S6K levels. (B) The intensity of the signal of the dots corresponding to p70S6K was quantified by densitometry and expressed relative to the control. (C) Western blotting detection of phosphorylated mTOR (S2448), total Akt and phosphorylated Akt (S473) in MCF-7 cells treated for 6 h with doxorubicin (1 μM) and/or melatonin (1 nM). (D) Western blotting detection of S6 phosphorylation (S240/S244) in MCF-7 cells treated for 6 h with doxorubicin (1 μM) and/or melatonin (1 nM).

Fig 4: The mechanism of how taurine relieves S. uberis infection by activating autophagy. From the above results, we infer that taurine enters the cells through transporters (TauT and PAT 1) to improve PTEN activity, promote the conversion of PIP3 to PIP2, and inhibit PI3K/Akt/mTOR signaling pathway, thereby attenuating the phosphorylation of ULK1 and ATG13 by mTOR, promoting the formation of ULK1-ATG13-FIP200 complex, and activating autophagy of MAC-T cells. S. uberis is recognized by both TLR2 and TLR4, internalizes into cells and may be ubiquitinated. Then, it is mediated by related receptor proteins into autophagosomes and fused with lysosomes to form autolysates. Finally, S. uberis is degraded and eliminated by proteases and hydrolases, which attenuates the stimulation to cells and reduces the activation of NF-κB, inhibits the secretion of inflammatory factors (such as TNF-α, IL-1β, and IL-6), and alleviates the excessive inflammatory response and damage caused by S. uberis infection.

Fig 5: PKCλ and PKMζ protein localize to central neurons in the dorsal horn and are increased by BDNF treatment. (A) Slices were prepared from mice and stained with an atypical PKC (aPKC) antibody (red) and the neuronal marker (found in soma and post-synaptic sites) SAP-102 (green). The top panel shows that aPKC immunoreactivity is localized primarily to the spinal dorsal horn. Lower panels (from inset) show that aPKC immunoreactivity is localized almost exclusively to neurons in the dorsal horn. (B) qPCR was used to assess mRNA expression of aPKC isoforms in spinal cord and kidney (N = 3). Spinal SNSs were prepared and mRNA or protein was isolated. Spinal SNSs were enriched for GluN1 mRNA and were βIII tubulin poor, they were likewise enriched in PSD95 protein as shown by Western blotting compared to equal protein concentration of whole spinal cord homogenate (input). C) SNSs were isolated from mouse lumbar spinal cord and exposed to increasing concentrations of BDNF for 15 min. BDNF dose-dependently increased mTOR, AKT and p70S6 kinase (p70) phosphorylation when standardized to total protein levels. BDNF also increased total levels of PKCλ PKMζ and CaMKIIα when compared to loading control βIII tubulin (N = 6). * p < 0.05, *** p < 0.001, one way ANOVA with Bonferroni post hoc test compared to Veh.