Research Paper Volume 13, Issue 13 pp 17097—17117

Figure 5. Neratinib and AR12 combine to reduce the expression of HSP90, HSP70, GRP78 and HSP27 via autophagy. Upper: Vero cells were treated with vehicle control, AR12 (2 μM), neratinib (50 nM) or the drugs in combination for 6h. Cells were fixed in place and in cell immunostaining performed to determine chaperone expression using antibodies whose epitopes are localized at the NH2- and COOH-termini of each protein (n = 3 +/-SD) * p < 0.05 less than vehicle control; ** p < 0.05 less than neratinib single agent value; ¶ p < 0.05 less than value determined using the COOH-terminal epitope antibody. Lower: HCT116 ATG16L1 T300 cells were transfected with a scrambled siRNA or with siRNA molecules to knock down the expression of Beclin1 or ATG5. After 24h, cells were treated with vehicle control, AR12 (2 μM), neratinib (50 nM) or the drugs in combination for 6h. Cells were fixed in place and in cell immunostaining performed to determine chaperone expression using antibodies whose epitopes is at the COOH-termini of each protein (n = 3 +/-SD) * p < 0.05 less than vehicle control; ** p < 0.05 less than neratinib single agent value.