Figure 2. JAM-A is elevated in CSCs.

Confocal micrographs show that JAM-A is co-expressed with CSC markers Sox2 (A) and integrin α6 (B) on GBM cells within an intracranial xenograft (T4121) using antibodies against JAM-A (green) and co-stained with Sox2 (red) or integrin α6 (red). Areas of interest indicated by yellow arrow and shown at higher power, and scale bar represents XX um JAM-A is elevated in CSCs (enriched by cell surface expression of CD133 from GBM xenograft tumor T4121) as compared with matched non-CSCs as shown in representative immunoblots that show separation between populations with respect to Sox2 and Olig2 (C). Bar graphs summarizing flow cytometry analyses between parental cells and CSCs demonstrates that CSCs have significantly higher expression of JAM-A as compared to parental unenriched tumor cells from GBM xenografts (T3691, T387, T4121, T4302, (D). CSCs grown in sphere culture (from tumor T4121) co-express JAM-A (green) and integrin α6 (red) as shown in confocal micrographs. An area of interest indicated by white box is shown with enlarged images below, nuclei were counterstained with Hoechst 33342, and scale bar represents 50 µm (E). Upon CSC differentiation (enriched from GBM xenograft tumor T4121) using a 10% FBS differentiation paradigm over a 7 day period, JAM-A expression is lost with concomitaant loss in CSC markers Sox2 and Olig2 and an increase in differentiation markers GFAP and Map2 as shown in representative immunoblots (F). Data for bar graphs displayed as mean values +/− standard deviation, blue value represents fold change, and ***p<0.001 as assessed by one-way ANOVA. See also Figure S2.