Fig 2: Biejiajian pill (BJJP) inhibited the growth and EMT of HCC by suppressing activation of the Akt/GSK-3β/Snail pathway in vivo. Mice were randomly grouped into six arms (n = 6): Normal Control arm (NC), Model arm (X), BJJP-low dose arm (L), BJJP-medium dose arm (M), BJJP-high dose arm (H), and sorafenib positive control arm (P). (A,B): The expressions of Akt, p-Akt (Ser 473), GSK-3β, p-GSK-3β (Ser 9), Cyclin D1, and PCNA in mice tumor tissues were examined using western blotting analysis. Representative western blots (A) and quantitative analysis of protein expression using ImageJ software (B) are shown. (C,D): The expressions of Snail, E-cadherin, N-cadherin, Vimentin, MMP-9 and VEGFA in mice tumor tissues were examined using western blotting analysis. Representative western blots (C) and quantitative analysis of protein expression using ImageJ software (D) are shown. Protein expression data were normalized to the GAPDH control and are shown as mean ± SD (n = 3). *, p < 0.05; **, p < 0.01 relative to the X arm (tumor model arm). One-way ANOVA was used to determine statistical significance. (E): IHC showed the inhibition of p-Akt (Ser 473), p-GSK-3β (Ser 9), Snail, N-cadherin and Cyclin D1, and promotion of E-cadherin levels in BJJP treated mouse tissues, compared with the tumor model arm (X) (scale bar, 100 μm, 200×).

Fig 3: SRC-mediated macrophage activation via activating PI3K/AKT signaling. (A,B) Western blot analysis of SRC expression in RAW264.7 and THP-1 cells. The plot gives a quantitative gray-scale analysis of the blot (n = 4). (C,D) Western blot analysis of the M1 marker NOS2 and IL-12 and the M2 marker ARG1 and IL-10 in RAW264.7 cells. The plot gives a quantitative gray-scale analysis of the blot (n = 3). (E,F) Western blot analysis of the M1 marker NOS2 and IL-12 and the M2 marker ARG1 and IL-10 in THP-1 cells. The plot gives a quantitative gray-scale analysis of the blot (n = 3). (G) Representative images of immunocytochemical staining analysis of NOS2 and ARG1 levels in RAW264.7 cells. Scale bar = 20 μm. (H,I) Western blot analysis of P-PI3K, P-AKT, and NF-κB in RAW264.7 cells. The plot gives a quantitative gray-scale analysis of the blot (n = 3). (J,K) Western blot analysis of P-PI3K, P-AKT, and NF-κB levels in THP-1 cells. The plot gives a quantitative gray-scale analysis of the blot (n = 3). All graphical data are the means ± SEMs from at least three independent experiments, each performed in triplicate. *P < 0.5 and **P < 0.01—levels of significance that are different from the control group.

Fig 4: Schematic diagram showing the mechanisms through which lncGm16410 mediates PM2.5-induced macrophages activation. When macrophage is exposed to PM2.5, lncGm16410 suppresses the regulation of SRC expression, leading to a subsequent increase in the latter. The interactions among lncGm16410, SRC, and PI3K/AKT will dictate the balance of M1/M2 macrophages. Macrophage apoptosis and activation result in the overproduction of inflammatory factors and their subsequent release into the lung. This process, in turn, causes inflammation in the lung.

Fig 5: 1,25(OH)2D3 alleviates the inhibition of H2O2 on the PI3K/Akt pathway. (a) Immunofluorescence staining of Akt and p-Akt in nucleus pulposus-derived mesenchymal stem cells (NPMSCs) is observed by a laser scanning confocal microscope (scale bar = 25 μm). (b) Quantitative analysis of the fluorescence expressions of Akt and p-Akt. (c, d) The protein expression and quantitative analysis of Akt and p-Akt in the different groups. All data are the mean ± SD. ∗P < 0.05 and ∗∗P < 0.01 compared with the control group; #P < 0.05 and ##P < 0.01 compared with the H2O2 group; &P < 0.05 and &&P < 0.01 compared with the 1,25(OH)2D3 group. LY: LY294002 group.