3.4 Confocal imaging shows that combination treatment
induces changes in Cx43 expression and intracellular distribution.
To better understand the mechanism of αCT1 enhanced cell killing, we
repeated the treatment in A172, U87, and BT169 cell lines using a
fluorescence conjugated αCT1 (FITC-αCT1) combined with TMZ. The
interaction between FITC-αCT1 and N-terminal Cx43 was visualized under
the confocal microscope. The complete procedure can be found in sections
2.6 and 2.7.
As shown in Figure 5 , Cx43 level did not show significant
changes after the TMZ (100 μM), or FITC-αCT1 (100 μM) treatment in any
of the three cell lines. However, in A172 and BT169 cell lines
(Figure 5A and 5B ), the combination of 100 μM TMZ and 100 μM
FITC-αCT1 treatment group significantly increased the number of Cx43
aggregates as well as the Cx43 signal intensity compared to the U87 cell
line, which only exhibits a slight increase in the Cx43 aggregates
number within the cell bodies (Figure 5C ). This observation
indicates that FITC-αCT1 combined with TMZ may induce changes in TMZ-
related signaling pathways and affect Cx43 protein functions and
activities in GBM cells.