Cord Blood Stem Cell-Mediated Induction of Apoptosis in Glioma Downregulates X-Linked Inhibitor of Apoptosis Protein (XIAP)
Figure 6
Upregulation of caspase-3 and Bax in nude mice brains.
Nude mice with pre-established intracranial human glioma tumors (U251 or 5310) were treated with hUCBSC by intracranial injection (2×105). Fourteen days after hUCBSC administration, the brains were harvested, sectioned and immunoprobed for caspase 3 using appropriate HRP-conjugated secondary antibody. (A) Expression of cleaved caspase-3 activity in brain sections. Bar = 200 µm. (B) Immunoblot analysis of proteins showing expression of cleaved caspase 3, TNF-alpha, TRAIL and cleaved PARP in tissue lysates of brains. (C) Quantitative estimation of (B). 40 µg of proteins were loaded onto 12–14% gels and transferred onto nitrocellulose membranes and probed with respective antibodies. Immunoreactive bands were visualized using chemiluminescence ECL Western blotting detection reagents and the reaction was detected using Hyperfilm-MP autoradiography film. (D) In vivo expression of Bax. Brain sections were immunoprobed for Bax and CD81 using appropriate secondary antibodies. Bax is labeled with Alexa Fluor-594 conjugated secondary antibody and CD81 is labeled with Alexa Fluor-488 conjugated secondary antibody. Bar = 200 µm. (E) Immunoblot analysis of proteins showing expression of Bax and Bad in tissue lysates of brains. 40 µg of proteins were loaded onto 14% gels and transferred onto nitrocellulose membranes and probed with respective antibodies. Immunoreactive bands were visualized using chemiluminescence ECL Western blotting detection reagents and the reaction was detected using Hyperfilm-MP autoradiography film. (F) Quantitative estimation of (E).