Fig. 3. The TLR4-Wnt5a signaling axis in lung epithelial tissues promotes pulmonary metastatic progression.

(A) Top: Primary BRAF^v600EPTEN^−/− tumor weight measurements after resection from TLR4^f/f (TLR4^+/+) and SPC-TLR4^−/− (TLR4^−/−) transgenic mice (n = 3). Bottom: Representative TRP2 IHC and TRP2 qrt-PCR analysis of lung tissues resected from TLR4^f/f and SPC-TLR4^−/− transgenic mice bearing BRAF^V600EPTEN^−/− tumors (n = 3). The IHC images are shown at 40×. Scale bars, 20 μm. ns, not significant. (B) Left: Representative low-magnification imaging of lungs resected from TLR4^+/+ and SPC-TLR4^−/− transgenic mice after BRAF^V600EPTEN^−/− tumor tail vein injection. Images are shown at 4×; Scale bars, 2000 μm. Right: Quantification of the area of tumor burden per whole lung (n = 3). (C) Left: Representative S100β IHC of lung tissues resected from TLR4^+/+and SPC-TLR4^−/− transgenic mice after BRAF^V600EPTEN^−/− tumor tail vein injection. Images are shown at 40×; Scale bars, 20 μm. Inset images are shown at 10×. Right: S100β-positive cells enumerated per lung based on IHC microscopy. HPF, high-power field. Analysis performed at 40×. (n = 4). (D) Lung weight measurements after resection from TLR4^+/+ and SPC-TLR4^−/− transgenic mice ± primary BRAF^V600EPTEN^−/− tumor (n = 4). g, gram. Statistical analysis was performed by two-way ANOVA followed by Sidak’s multiple comparisons test. All two-group comparisons were analyzed using unpaired t tests. All data are representative of two to three independent experiments and expressed as mean values ± SEM. ns, nonsignificant; *P < 0.05 and ***P < 0.0005.