Fig. 4: LAPTM5 blocks BMP signal and negatively regulates BMPR1A.

a Differentially expressed cancer-related gene sets (C2_PATHWAYS) with high LAPTM5 expression in the TCGA KIRC cohort and KIRP cohort (Top 5). NES, normalized enrichment score. FDR q, false discovery rate q value. GSEA output of genes in the PID_BMP_Pathway by LAPTM5 high and low expression groups from the KIRC cohort (b) and KIRP cohort (c) in the TCGA database. ES, enrichment score. d IB analysis of Renca^luci parental and derivative cells treated with Mock (ddH₂O), or recombinant murine BMP4 (10 ng/mL or 20 ng/mL) for 60 min. e IHC analysis of p-Smad 1/5/8 levels in lung metastases of Renca^luci. Scale bar, 50 μm. f IB analysis of control and Laptm5-overexpressing Renca cells (left panel) or control and Laptm5-silenced Renca^LuM2b cells (middle and right panel) treated with BMP4 (10 ng/mL) for indicated times. g Representative images of tumor sphere assay of control and Laptm5-overexpressing Renca cells treated with or without BMP4 (30 ng/mL) for 7 days, and quantification of tumor sphere formation of indicated groups (n = 3 per group). Scale bar, 200 μm. h Representative images of tumor sphere assay of control and Laptm5-silenced Renca^LuM2b cells treated with or without DMH1 (5 μM) for 7 days, and quantification of tumor sphere formation of indicated groups (n = 3 per group). Scale bar, 200 μm. i qRT-PCR analysis of stemness markers in indicated cells. n = 3 per group. j Semiquantitative RT-PCR analysis of BMP receptors in control and Laptm5-overexpressing Renca^luci cells with Actb as control. k IB analysis of BMPR1A in control and Laptm5-overexpressing Renca cells treated with BMP4 (10 ng/mL) for indicated times. l Ratio of BMPR1A protein remaining in control and Laptm5-overexpressing Renca cells treated as in (k) (n = 3 per group). m qRT-PCR analysis of Bmpr1a in control and Laptm5-overexpressing Renca cells treated with BMP4 (10 ng/mL) for indicated times. n = 3 per group. Immunoblots are representative of three biological replicates. Data in bar graphs are presented as mean ± SD. Two-tailed Student’s unpaired t-test were used for statistical analysis in all panels. Source data are provided as a Source data file.