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. 2015 Apr 2;6(11):8709–8721. doi: 10.18632/oncotarget.3325

Figure 3. CD44-depletion reduces both the expression and nuclear localization of the stemness factors, Nanog, Sox2, and Oct4.

Figure 3

MDA-MB-231 and MCF-7 cells were transfected with scrambled siRNA (scRNA) and CD44 siRNA, and then treated with 5 μM of GSI for 24 hr. (A) mRNA (upper) and protein levels (lower) of CD44 and stemness factors were detected with an RT-PCR and western blot analysis. (B and C) the changes in the localization of cleaved CD44ICD and stemness factors were detected using western blot analysis. The cells were treated with 5 μM of GSI for the indicated times. GAPDH and Lamin A/C were used as loading controls. (D) The changes in the localization of CD44ICD and its co-localization with stemness factors were detected during treatment with 2 μM of GSI for 12 hr in MDA-MB-231 and MCF-7 cells using an immunocytochemical analysis as described in “materials methods”.