Figure 5
Analyses of HA/CD44-mediated Bcl-2 and IAP expression in MDA-MB-468 cells. (A) Cell lysates from cells [treated with no HA (lane 1) or with HA (lane 2) or pretreated with anti-CD44 antibody plus HA (lane 3) or treated with non-immune IgG (without HA) (lane 4) or with HA (lane 5)] were immunoblotted with anti-BCL-2 antibody (a) or anti-cIAP-1 antibody (b) or anti-cIAP-2 antibody (c) or anti-XIAP antibody (d) or anti-actin antibody (e), respectively. [The ratio of Bcl2 (a) or cIAP1 (b) or cIAP2 (c) or XIAP (d) and actin (c) was determined by densitometry, and normalized to untreated (without HA treatment) value or non-immune IgG (no HA) (designated as 1.00)]. (B) Cell lysates from cells [transfected with scrambled siRNA with no HA (lane 1) or with HA (lane 2) or transfected with c-Jun siRNA plus HA (lane 3)] were immunoblotted with anti-BCL-2 antibody (a) or anti-cIAP-1 antibody (b) or anti-cIAP-2 antibody (c) or anti-XIAP antibody (d) or anti-actin antibody (e), respectively. [The ratio of Bcl2 (a) or cIAP1 (b) or cIAP2 (c) or XIAP (d) and actin (e) was determined by densitometry, and normalized to the value of scrambled siRNA-treated samples (no HA) (designated as 1.00)]. (C) Cell lysates from cells [transfected with negative miRNA control plus no HA (lane 1) or with HA (lane 2) or transfected with anti-miRNA-21 plus HA (lane 3) or treated with JNK inhibitor plus HA (lane 4)] were immunoblotted with anti-cIAP-1 antibody (a) or anti-cIAP-2 antibody (b) or anti-XIAP antibody (c) or anti-actin antibody (e), respectively. [The ratio of Bcl2 (a) or cIAP1 (b) or cIAP2 (c) or XIAP (d) and actin (e) was determined by densitometry, and normalized to the value of negative control miRNA-treated samples (no HA) (designated as 1.00)]. The values expressed represent an average of triplicate determination of 4 experiments with an SD of less than 5%.