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. 2015 Oct 22;6(42):44452–44465. doi: 10.18632/oncotarget.6090

Figure 4. Nuclear CD44 participated in transcriptional regulation of C3A-iCSCs.

Figure 4

A. Immunofluorescence staining of CD44 in C3A cells, C3A-iCSCs and U87 cells. U87 cells were used as a positive control and C3A cells as a negative control. Red indicated positive staining. Nuclei were counterstained with Hoechst 33342 (blue). Scale bar, 40 μm. B. Immunofluorescence staining of CD44 expression in C3A-iCSCs and C3A-iCSCs cultured in DMEM containing 10% fetal bovine serum medium for 24 and 48 h. Red indicated positive staining. Nuclei were counterstained with Hoechst 33342 (blue). Scale bar, 40 μm. C. ChIP-qPCR using an anti-CD44 antibody and anti-IgG control in C3A cells and C3A-iCSCs. The CYCLIN D1 promoter was used as a positive control which have been reported that CD44 protein could bound to [17]. SOX2, c-MYC and CYCLIN D1 represented respectively their promoters. Samples were analyzed by qPCR. Error bars showed the standard deviation of three independent ChIP-qPCR assays. D. Luciferase activity assay about CD44 transcriptional regulation ability. Luciferase reporter plasmids pGL3 c-MYC promoter and pGL3 SOX2 promoter were co-transfected with the pCMV3-CD44 plasmid and Renilla control vector in 293T cells. Luciferase activities were measured after 48 h and presented as relative to the activity of Renilla luciferase. pGL3 SOX2 promoter and pGL3 c-MYC promoter groups were control.