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Fig. 4 | BMC Cancer

Fig. 4

From: LncRNA MCF2L-AS1 aggravates the malignant development of colorectal cancer via targeting miR-105-5p/RAB22A axis

Fig. 4

RAB22A is a target of miR-105-5p and participates in CRC progression. A. Bioinformatics tools (miRmap, microT and RNA22) were used to determine the potential target genes of miR-105-5p. B. The expression of candidate mRNAs was assessed via RT-qPCR analysis after up-regulating miR-105-5p. C. RAB22A expression was tested by RT-qPCR after down-regulating MCF2L-AS1. D. The expression of RAB22A was examined in CRC cell lines and human normal colon epithelial NCM-460 cell line. E. RIP assay was conducted to test the binding relationship among MCF2L-AS1, RAB22A and miR-105-5p. F. RNA pull down assay was carried out to test the connection between miR-105-5p and RAB22A. G. The binding sites between miR-105-5p and RAB22A were predicted by ENCORI website. H. The binding correlation between miR-105-5p and RAB22A was testified through luciferase reporter assay. I. The interplay among MCF2L-AS1, RAB22A and miR-105-5p was tested by RT-qPCR assay. J. The interference efficiency of RAB22A was tested via RT-qPCR assay. K and L. IF and EdU assays were adopted to assess the cell proliferation ability after RAB22A was silenced. M and N. Transwell assay was performed to evaluate cell migration and invasion upon RAB22A silencing. O and P. Flow cytometry assay and caspase-3/8/9 activity analysis were adopted to assess cell apoptosis in CRC under different transfection conditions when RAB22A was silenced. Q. The EMT process was detected through IF assay after RAB22A was knocked down in different groups. Adjustments of individual color channels were made on ‘Merge’ figures. The statistical analysis of Fig. 4H was tested with two-way ANOVA, and the statistical analysis of Fig. 4B-F, I-Q was tested with one-way ANOVA. *P < 0.05, **P < 0.01

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