Extended Data Fig. 4: LTβ promotes bone metastasis.
From: Lymphotoxin-β promotes breast cancer bone metastasis colonization and osteolytic outgrowth
a-b, The mRNA expression levels of LTB (a) and ADGRF5 (b) in SCP28 cells with LTβ- (a) or ADGRF5- (b) OE were determined by qPCR. n = 3 biologically independent samples. c, 103 SCP28 cells with -Vector control, LTβ-OE, or ADGRF5-OE were seeded into 96 well plate for cell culture. Relative cell number was monitored by in vitro luciferase assay. n = 3 biologically independent samples. d, 105 SCP28 cells with -Vector control, LTβ-OE, or ADGRF5-OE were IC injected into female nude mice. Bone metastasis burden was monitored by BLI imaging. n = 5, 7 or 7 for Vector control, LTβ group, or ADGRF5 group. e, Representative BLI, μCT, X-ray, H&E staining, and TRAP staining images of mice from experiment in (d). Scale bar = 50 μm. BLI scale: x 107 p/sec/cm2/sr. f, Quantification of the number of TRAP+ osteoclasts from experiment performed in (e). n = 4 biologically independent samples. g, Quantification of osteolytic lesion areas based on X-ray images from experiment performed in (e). Group size as in (d). h-i, LTβ (h) and ADGRF5 (i) in SCP28 cells were knocked down by shRNA constructs, mRNA levels were determined by qPCR. n = 3 biologically independent samples. j, SCP28 cells with -Vector control or LTβ-OE were injected in the mammary fat pads of female nude mice. Tumor volume was measured weekly. n = 5 mice. k, 105 4T1 cells with -Vector control or LTβ-OE were tail-vein injected into 6-week-old female BALB/c mice. Lung metastasis burden was monitored by BLI imaging. n = 8 mice. l, Representative BLI, lung image, and H&E staining of mice from experiment performed in (k). Scale bar = 5 mm. BLI scales: x107 p/sec/cm2/sr. m, Quantification of lung metastatic nodules from (k). n = 8 lungs per group. Arrows indicate the osteolytic bone lesion areas (e). B, bone tissue area; T, tumor area (e). Data presented as mean ± SEM (c,f,g,j,k) or SD (a,b,d,h,i,m). The P values were determined by two-way repeated measures ANOVA (c,d,j,k) or two-tailed unpaired t-test (a,b,f,g,h,i,m).
