Journal: BMC Cancer

Article Title: HSP90 inhibition sensitizes head and neck cancer to platin-based chemoradiotherapy by modulation of the DNA damage response resulting in chromosomal fragmentation

doi: 10.1186/s12885-017-3084-0

Figure Lengend Snippet: AUY922 abrogates ATR-CHK1 signaling allowing increased chromosomal fragmentation in response to CCRT. a Scheduling showing 0 h time point post 16 h AUY922 addition but pre-RT, cisplatin or combined CCRT addition and subsequent time point analysis post as used in panels b - f. b AUY922 disruption of ATR-CHK1 signaling in response to CCRT alongside depletion of total RAD51. c Mitotic accumulation as measured by FACS analysis of phospho-histone H3 positive cells. d Co-staining for phospho-histone H3 and γH2Ax was analyzed by FACS. Population plotted is the percentage of the total cell number positive for both high γH2Ax levels and the mitotic marker phospho-histone H3. e γH2Ax staining in mitotic cells was confirmed in HNSCC cell lines by confocal microscopy, DAPI as nuclear stain. Nuclei with mitotic morphology indicated by arrows. f Micronuclei quantification of DAPI stained confocal images at 24 h in response to CCRT and AUY922. Values are mean ± SEM of at least three independent experiments. Statistical analysis by 2-tailed t -test; * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Cells were stained for mitosis or DNA double-stranded breaks with rabbit anti-phospho-histone H3 S10 (DD2C8) AlexaFluor647 or anti-phospho-histone H2Ax S139 (20E3) AlexaFluor488 (Cell Signaling, MA, USA) using the manufacturer’s protocol.

Techniques: Disruption, Staining, Marker, Confocal Microscopy