Dose-dependent growth inhibition in response to 72h exposure to lapatinib (a), canertinib (b), or afatinib (c) on malignant human cell lines with no HER2 expression (Jurkat), with moderate HER2 expression (T47D) and with HER2 overexpression (SKBR3).

<p>Means ± SD, n = 3.</p

Schematic representation of HER-dependent signal transduction pathways associated with cell proliferation and apoptosis.

<p>Schematic representation of HER-dependent signal transduction pathways associated with cell proliferation and apoptosis.</p

Effect of the HER-TKI canertinib on the expression of apoptosis-related PARP-1 cleavage products as demonstrated by Western blot analysis of T47D cells grown for 48h (lanes 1–3) and 72h (lanes 4–6) in the absence (lanes 1 and 4), or in the presence of 1μM (lanes 2 and 5) and 10 μM (lanes 3 and 6) canertinib.

<p>Actin was probed on a separate membrane to roughly estimate protein loading.</p

Dose-dependent growth inhibition in response to 72h exposure to the JNK inhibitor SP600125 (a) or the STAT5 Inhibitor II IQDMA (b) on SKBR3 and T47D human breast cancer cells.

<p>Means ± SD.</p

Effect of canertinib (white bar), lapatinib (light grey bar) and afatinib (dark grey bar) on activation (phosphorylation) of ERK/MAPK1/2, STAT3, CREB, STAT5A/B, p70 S6 kinase, IkBa, JNK, and p38 in SKBR3, T47D and Jurkat cells.

<p>Serum-depleted cells were exposed for 6 h to vehicle (0.1% DMSO) or to 1 μM of the TKIs followed by a 3-min challenge with 100 ng/ml EGF and 1 nM HRGb1 and subjected to Milliplex assays. Results are shown relative to vehicle control (100%). Means ± SD.</p

Dose-dependent effect of SP600125 when given for 72h (a) and time-dependent effect of 5μM canertinib after 2 and 6 hours (b) on apoptosis as examined by Milliplex immunoassay for detection of active (cleaved) caspase 3 in SKBR3 and T47D cells.

<p>Results are shown relative to vehicle (0.1% DMSO) control. Means ± SD.</p

Antiproliferative effect of ZA in the absence or presence of different concentrations of estradiol.

<p>Antiproliferative effect of rising concentrations of ZA (0, 6.25 and 12.5μM) in the three cell lines MCF-7 (A), T47D (B) and MDA-MB-231 (C) in the absence (black bars) or the presence of 1nM (grey bars) and 10nM (white bars) estradiol (EZ4U).</p

Dose-dependent antiproliferative effect of ZA.

<p>Dose-dependent antiproliferative effect of ZA on the three cell lines in the absence (A) and the presence (B) of 1nM estradiol (EZ4U). Abbreviations: OD = optical density.</p

Pro-apoptotic effect of ZA measured by Annexin/PI staining.

<p>Pro-apoptotic effect of rising concentrations of ZA in the three cell lines MCF-7 (A), T47D (B), MDA-MB-231 (C) in the absence or the presence of 1nM estradiol measured by AnnexinV/PI staining.</p

Proliferative effect of estradiol.

<p>Proliferative effect of 0, 1 and 10nM estradiol on the three cell lines MCF-7 (black), T74D (grey) and MDA-MD-231 (white) in the absence of ZA (EZ4U). Abbreviations: OD = optical density.</p