Fig. 2: LAPTM5 mediates lung-specific metastasis.

a Principal component analysis (PCA) of Renca^luci parental and derivative cells. b Overlap of up-regulated genes in Renca^LuM2b cells (LuM) as compared to Renca^Parental (Par), Renca^BrM2b (BrM) and Renca^BoM2 (BoM) cells (left panel) and further overlap with up-regulated genes in RCC lung metastases (L-Mets) as compared with primary RCC without lung metastases (pri-RCC) in the Jon_Renal_Cancer dataset (right panel). c Heatmap showing the mRNA expression of Ctss, Laptm5 and Igfbp5 in Renca^luci parental and derivative cells measured by qRT-PCR, relative gene expression in each cell was compared with parental-1 (Par-1). d Immunoblotting (IB) analyses of LAPTM5 expression in Renca^luci parental and derivative cells. e Immunohistochemistry (IHC) analysis of LAPTM5 staining in RCC in situ (renal subcapsular injection) and lung, bone, brain metastases. HE staining was used to show the site of bone metastasis. T, tumor. Scale bar, 200 μm. f IB analysis of control and Laptm5-overexpressing Renca^luci cells or control and Laptm5-silenced Renca^LuM2b cells. g Schematic illustration for intracardiac cell inoculation and metastasis detection. Representative bioluminescent images of lung metastases (h), brain and bone metastases (i) in mice treated as in (g) with Renca^luci and derivatives. j Representative X-ray images of bone metastases in mice treated as in (g). k Quantification of photon flux changes of lung metastases in (h) (n = 7 mice per group). l, m Representative bioluminescent images of lung metastases (l), brain and bone metastases (m) in mice treated as in (g) with Renca^LuM2b and derivatives. n Representative X-ray images of bone metastases in mice treated as in (g). o Quantification of photon flux changes of lung metastases in (L) (n = 7 mice per group). p Quantification of photon flux of brain and bone metastases at 15 days after I.C. inoculation with indicated cells (n = 7 mice per group). Boxes represent data within the 25th to 75th percentiles, n.s., not significant. Immunoblots are representative of three biological replicates. In k and o, the data are presented as mean ± SEM, in (p), the data are presented as whisker plots: midline, median; box, 25–75th percentile; whisker, minimum to maximum values. Two-tailed Student’s unpaired t-test were used for statistical analysis in all panels. Source data are provided as a Source data file.