Fig 2: A schematic diagram of model displaying the implication of MIIP in PKCε activation-induced tumor metastasis. PKCε phosphorylates MIIP at Ser303 and promotes its interaction with RelA, which protects RelA from deacetylation by HDAC6; in turn, this leads to enhancement of RelA transcriptional activity and increase of genes expression relevant to cell invasion. PP1 acts as a phosphotase being able to dephosphorylate MIIP phosphorylation by PKCε. In cancer cells, PKCε activity is hyperactive while PP1 protein levels is downregulated; thereby, PKCε /MIIP/RelA signaling is reinforced and faciliates tumor metastasis

Fig 3: PKCε phosphorylated MIIP and promoted MIIP–RelA interaction. a HCT116 cells were treated with or without EGF. Cellular extracts subjected to immunoprecipitation with an anti-Flag antibody. The immunoprecipitates were treated with CIP (10 units), followed by immunoblotting analysis. b, c In vitro phosphorylation analyses were performed by mixing the purified active PKCε with the indicated purified GST-MIIP proteins in the presence of [γ-32P]ATP. Ser303 of MIIP is evolutionarily conserved in the indicated species (c, left panel). d HCT116 cells expressed with WT MIIP or MIIP S303A were treated with or without EGF for 30 min. e HCT116 cells expressed with WT MIIP or MIIP S303A were treated with or without EGF (100 ng/ml) for 10 h. f HCT116 cells with depletion of MIIP, and reconstituted expression of WT rMIIP or rMIIP S303A were treated with or without EGF (100 ng/ml) for 10 h. g HCT116 cells with depletion of MIIP, and reconstituted expression of WT rMIIP or rMIIP S303A were treated with or without EGF for 10 h. Relative mRNA levels were analyzed by q-PCR. h HCT116 cells with depletion of MIIP, and reconstituted expression of WT rMIIP or rMIIP S303A were treated with or without EGF (100 ng/ml). Cell invasion assays were performed. In e, f, ChIP analyses with indicated antibodies were performed. The primers covering RelA binding site of MMP2 gene promoter region were used for the q-PCR. The Y axis shows the value normalized to the input. In a–d, immunoblotting analyses were performed using the indicated antibodies. In e–h, the values are presented as mean ± s.e.m. (n = 3 independent experiments), ** represents P < 0.01 (Student’s t-test) between the indicated groups

Fig 4: Intervention of exercise and leucine enhanced adiponectin secretion and reduced adipose tissue inflammation under HFD. C57BL/6J male mice of 5–6 weeks were fed with LFD or HFD. Leucine with a concentration of 1.5% and/or moderate intensity exercise was added in the 10th week and administrated for 4 weeks. (A) Adipose tissue immunohistochemistry. The sections of adipose tissue were immunostained for F4/80. For (B, C), the levels and phosphorylation states of NF-?B p65 were determined using western blots. For (D, E), quantification of inflammatory signaling (arbitrary units) for (B, C), respectively. For (F, G), changes in adipose mRNA level of adiponectin by RT-PCR under LFD and HFD, respectively. The data are mean ± s.d. (error bars), n = 6 mice per group. *p < 0.05, HFD versus HFD + Ex + Leu.

Fig 5: Combined treatment with RFA and MLT reduced tumor malignancy in non-ablated areas, altered tumor metabolism, and the tumor microenvironment. a The integrative multi-omics analysis of cancer proliferation and malignancy-related pathways. Heatmap of alteration pathways in the transcriptomic and proteomic subtypes. Left, alteration pathways identified from the transcriptome. Right, alteration pathways are identified from the proteome. Color of each cell represents the average ssGSEA enrichment scores of that subtype; red denotes activation and blue denotes inhibition. Blank cells represent non-enrichment. b The validation of cancer proliferation and malignancy-related pathways. Western blot analysis of P53 (Cleaved-BID and P53), Wnt (C-myc, β-catenin, P-β-catenin, and Cyclin D1), Hedgehog (β-Arrestin 1 and Gli1), MAPK (P-erk, Foxo3, PKC, ERK2, p-STAT3 and STAT3), and NFкB (Rela, BTK, CSNK2A1) pathway-altered proteins in the RFA + MLT, RFA, MLT, and Control groups; The normalized altered gene expression of c P53 (Mdm4, Siva1, Fas, and Igfbp3), d NFкB (Ptgs2, Cxcl1, and Il1b), e Wnt (Sox17), and f MAPK (Hspa1b, Pgf, Fas, Angpt2, Cacna1g, Kit, Hspa1a, Map2k6, Dusp4, and Il1b) signal pathways in the RFA + MLT, RFA, MLT, and Control groups (*P < 0.05; **P < 0 .01; ***P < 0. 001)