Fig. 5

HA-CPNs Downregulate the Expression Levels and Signaling Activity of CD44 in GBM. (A) U-251 MG cells were treated with HA-CPNs and/or vehicle control (HA-CPN vehicle) and subjected to flowcytometry analysis of the indicated TIC populations; (B) Assessment of the protein expression levels of CD44 and CD44 intracytoplasmic domain (CD44-ICD) via Western blotting in FACS derived HA-CPN + and HA-CPN- cells (top). Protein levels quantified using densitometry and shown as ratio to Actin in Integrated Density Values (IDV) (bottom, graphs); (C) mRNA expression of signature markers of diverse endocytotic mechanisms in U-251 MG cells treated with 24 and/or 48 HA-CPNs/cell relative to HA-CPN vehicle control (dotted line); (D) CD44 and CD44-ICD protein expression levels in U-251 MG cells treated with HA-CPNs and/or HA-CPN vehicle control in the presence of Endosidin 9 or vehicle control (left). Protein levels quantified using densitometry and shown as ratio to Actin in Integrated Density Values (IDV) (right); (E) Flowcytometry analysis of CD44 expression levels in the indicated GBM patient samples treated with HA-CPNs and/or HA-CPN vehicle control. Data shown as mean ± s.d, n = 3, *p < 0.05, **p < 0.01, ***p < 0.001; Student’s t-test. Original blotted membranes of Western can be found in Fig. S7.