Fig 2: Perioperative ibuprofen intervention inhibited the activation of tau phosphorylation related kinases and cellular stress signaling pathways induced by laparotomy. a & b Effect of ibuprofen on GSK3ß and phosphatase (PP2A) in the frontal cortex (a) and hippocampus (b), c & d stress (Jak2/Stat3 and JNK) and cell survival (ERK) signaling pathways in the frontal cortex (c) and hippocampus (d). For each panel, n = 6, *p < 0.05, **p < 0.01, ***p < 0.001

Fig 3: Production of cytokines and activation of NF-?B and STAT3 are enhanced in Mettl3-null macrophages co-cultured with tumour cells.a qRT-PCR analysis of Il-6 expression levels in B16 and LLC cells incubated with WT or KO BMDM culture medium for 24 hours in the presence or absence of BAY-11-7082 (5 µM) (n = 5 mice per group). b B16 and LLC cells were treated with TNF-a (40 ng/mL) or TNF-a + BAY-11-7082 (5 µM), followed by qRT-PCR analysis of Il-6. n = 3 independent experiments. c B16 and LLC cells co-cultured with WT or KO BMDMs in the presence of BAY-11-7082 (5 µM) or an anti-TNF-a Ab (20 µg/mL) for 8 h. qRT-PCR analysis of Il-6 expression levels in WT or KO BMDMs was performed (n = 5 mice per group). d Western blot analysis of the indicated proteins in WT and KO BMDMs co-cultured with B16 cells in the presence or absence of the anti-TNF-a Ab (20 µg/mL). The blots are representative of n = 3 independent experiments. e qRT-PCR analysis of Il-6 expression in WT and KO BMDMs co-cultured with B16 or LLC cells in the presence or absence of an anti-IL-6 Ab (20 µg/mL) (n = 5 mice per group). f Western blot analysis of the indicated proteins in WT and KO BMDMs co-cultured with B16 cells in the presence or absence of the anti-IL-6 Ab (20 µg/mL). The blots are representative of n = 3 independent experiments. Data are shown as the mean ± SD. P values were determined by a two-tailed t-test (a–c and e). *P = 0.05, **P < 0.01 and ***P < 0.001. The source data are provided as a Source data file.

Fig 4: The arsenic-induced miR-21 increase and PDCD4 suppression contribute to malignant cell transformation.BEAS-2B cells were maintained in a medium containing various concentrations of arsenic (0.1, 0.25 and 0.5 µM) for 6 months. (A,B) Cells were cultured in 0.35% soft agar for 5 weeks and number of colonies in the entire dish counted. (A) Representative images of control (left panel) and arsenic-treated (right panel) colonies. (B) Colony number increased in a dose-dependent manner. (C) The relative miR-21 level, determined by Taqman real-time PCR, increased in a time- and dose-dependent manner. (D) Total cell lysates were prepared for western blot analysis after 2, 4 and 6 months exposure to arsenic using specific antibodies against PDCD4, p47phox, pSTAT3 and STAT3. Apparent protein levels for PDCD4 decreased and P47phox and pSTAT3 increased in a time- and dose-dependent manner. (E) Representative images of fluorescence immunostaining for PDCD4 and pSTAT3 after 2, 4 and 6 months exposure to arsenic, and confirm results from western blot analysis. Data presented in the bar graphs are the mean ± SD of three independent experiments. *Indicates a statistically significant difference compared to control with p < 0.05.

Fig 5: Arsenic-induced ROS generation and STAT3 activation are essential for the miR-21 increase and PDCD4 suppression.(A–D) BEAS-2B cells with stable overexpression of catalase or their corresponding vehicle vector were exposed to arsenic (0 or 0.5 µM) for 6 months. (A) Catalase overexpression was verified by western blot analysis. (B) The relative miR-21 level was determined by Taqman real-time PCR and decreased with catalase overexpression. (C) Cell lysates were prepared to determine the protein level of PDCD4 by western blot analysis. Apparent PDCD4 levels increased with catalase overexpression. (D) Anchorage-independent colony growth, assessed as previously described, decreased with catalase overexpression. (E–H) p47phox expression was stably knocked down in BEAS-2B cells and exposed to arsenic (0 or 0.5 µM) for 6 months. (E) P47phox knockdown was confirmed by western blot analysis. (F) Relative miR-21 level, determined by Taqman real-time PCR was decreased. (G) Cell lysates were prepared to evaluate PDCD4 protein levels by western blot analysis. An apparent restoration in PDCD4 levels was observed. (H) Anchorage-independent colony growth, assessed as previously described, was also decreased compared with respective vector controls. (I–K) BEAS-2B cells with stable knockdown of STAT3 or their corresponding vehicle vector were exposed to arsenic (0 or 0.5 µM) for 6 months. (I) The Relative miR-21 level, determined by Taqman real-time PCR, decreased compared with respective vector controls. (J) Cell lysates were prepared to determine the protein level of PDCD4 by western blot analysis. Apparent PDCD4 levels increased. (K) Anchorage-independent colony growth, assessed as previously described, decreased compared with respective vector controls. Data presented in the bar graphs are the mean ± SD of three independent experiments. *Indicates a statistically significant difference compared to control with p < 0.05.