Fig 1: FMNL1 expression in cell lines and FMNL1 knockdown inhibits proliferation, migration and invasion in ccRCC cells. Values were presented as the mean ± SD.*p<0.05, **p<0.01 and ***p<0.001. A) Western blot analysis of FMNL1 in normal kidney cell line (HK2) and renal cancer cell lines (786-0, Caki-1 and SNU-349). B) Western blot analysis of FMNL1 expression after 786-O and Caki-1 were infected by lentivirus. C) Colony formation assay of infected 786-O and Caki-1 cells with FMNL1 knockdown. D) Wound healing analysis of infected 786-O and Caki-1 cells with FMNL1 knockdown. E) Transwell analysis was performed to calculate the invasive cells in infected 786-O and Caki-1.
Fig 2: FMNL1 gene expression profiles and correlated with cell migration. A) FMNL1 gene expression profiles across all tumor samplesand paired normal tissues based on RNA-Seq data from TCGA database using GEPIA2. B) The correlation betwwen FMNL1 mRNA and cell migration and actin cytoskeleton organizaton were analyzed by GSEA assay. |NSE|>1, FDR<0.25,P<0.05 were considered as statistically significant.
Fig 3: Validation of FMNL1 expression by RT-qPCR, WB and IHC. A) Real-time PCR analysis was used to evaluate the expression level of FMNL1 mRNA in 10 paired ccRCC and adjacent normal tissues. Values were presented as the mean±SD.*p<0.05, **p<0.01. B) Densitometric analysis and quantification of Western blotting band were performed. The Box plots show the average concentration of FNML1 was higher in ccRCC than in the normal kidney tissues.β-actin was used as a loading control. C) IHC was perforemd on 81 tumor tissues and 16 normal tissues (Original magnification x 200).The sum Integrated Optical Density (IOD) of each IHC photo was calculated by Image-Pro Plue 6.0. P<0.05 was considered as statistically significant. D) Kaplan-Meier analysis presented the correlation between high expression of FMNL1 eand poorer overall survival in 52 ccRCC patients(p<0.0001, log-rank test).
Fig 4: Validation of 2 potential biomarkers from upregulated expression proteins. A) Hierarchical clustering analysis of 29 significantly up-regulated genes (FC>5, P<0.01) was performed on the Heml 1.0.3.7 derived from TCGA-KIRC datasets. The expression of 29 genes were consistent with our quantitative proteomic results. B) Kaplan-Meier curves for OS of patients with ccRCC were performed. Patients with ccRCC were divided into “high”and “low”groups based on median values of genes mRNA sequencing quantification data from TCGA-KIRC project. C) ROC curve analysis was performed to evaluate the specificity and sensitivity of the genes FMNL1 and CORO1A for differentiating ccRCC and paired normal tissues in the TCGA-KIRC datasets.
Supplier Page from Abcam for Anti-FMNL1 antibody