Fig 2: Inhibition of YAP1 activity ameliorates inflammation in ALI model mice. The lung tissues and BALF were collected at 72 h after LPS administration. (A) Immunofluorescence staining of the macrophage marker F4/80 (green) and the neutrophil marker MPO (red). Nuclei were counterstained with DAPI (blue). (B) Quantification of positive neutrophils. (C) Determination of MPO activity. The levels of IL‐10 (D), IL‐6 (E), and TNF‐α (F) were determined by ELISA assays. (G) Western blotting and densitometric quantification of TNF‐α, CD80, iNOS, CD206, and Arg1. All data are presented as the mean ± SD (n = 5). ns, no significance; *p < 0.05, **p < 0.01, ***p < 0.001 between two groups. MPO, myeloperoxidase; TNF‐α, tumor necrosis factor α; IL‐6, interleukin 6; IL‐10, interleukin 10. CD206, mannose receptor; iNOS, inducible nitric oxide synthase; CD80, cluster of differentiation 80; Arg1, Arginase 1.

Fig 3: Knockdown of RCN1 in Cal-27 and SCC-25 cells inhibits M2 polarization of THP-1 macrophages. (A) The TIMER database was used to analyse the correlation between M2 macrophage infiltration and RCN1 expression in OSCC. (B) RT‒qPCR was used to detect the expression level of the M0 macrophage marker CD68. (C) RT‒qPCR was used to detect the expression level of the M2 macrophage marker expression in cocultured TAMs. (D) Protein levels of CD206, IL-10, and ARG1 in cocultured TAMs. *p < 0.05, **p < 0.01, ***p < 0.001.

Fig 4: Inhibition of YAP1 activity promoted M2 polarization in LPS‐ and IL4‐induced BMMs. Cells were induced by 50 ng/mL LPS or 25 ng/mL IL4 for 24 h, and then treated with or without 2 μg/mL VER for 72 h. (A) Western blotting and densitometric quantification of CD80, iNOS, CD206, Arg1, p‐YAP1, YAP1, and CCL2. (B) Western blotting and densitometric quantification of p‐YAP1, YAP1 in the nucleus and cytoplasm. (C) The mRNA expression of Cd80, Nos2, Mrc1, Arg1, Yap1, and Ccl2 determined by qRT‐PCR. All data are presented as the mean ± SD (n = 5). ns, no significance; *p < 0.05, **p < 0.01, ***p < 0.001 compared with LPS‐treated group; # p < 0.05, ## p < 0.01, ### p < 0.001 compared with IL4‐treated group. CD206, gene symbol: Mrc1; iNOS, gene symbol: Nos2; CD80, gene symbol: Cd80; Arg1, gene symbol: Arg1; YAP1, Yes‐associated protein 1 (gene symbol: Yap1); CCL2, chemokine (C‐C motif) ligand 2 (gene symbol: Ccl2).

Fig 5: LATS1/YAP1 regulated the polarization state of M1/M2 macrophage in BMMs. Cells were transfected with siRNA‐YAP1 (si‐Yap1), siRNA‐LATS1 (si‐Lats1), or siRNA negative control (si‐NC). 72 h after siRNA transfection, the cells were collected for qRT‐PCR and western blotting assays. (A) Western blotting and densitometric quantification of p‐YAP1, YAP1, LATS1, CCL2, CD80, iNOS, CD206, and Arg1. (B) Western blotting and densitometric quantification of p‐YAP1, YAP1 in the nucleus and cytoplasm. (C) The mRNA expression of Yap1, Ccl2, Cd80, Nos2, Mrc1, and Arg1 determined by qRT‐PCR. All data are presented as the mean ± SD (n = 5). ns, no significance; *p < 0.05, **p < 0.01, ***p < 0.001 between two groups. LATS1, large tumor suppressor 1 (gene symbol: Lats1).