Data are presented as mean ± SD *p < 0.05 **p < 0.005 of triplicate repeats. C Clonogenic assays were performed at 9 days post-treatment with dose-dependent olaparib (0, 1, 5 µM) in the indicated cell lines. All experiments in each cell line were repeated at least 5 times. Statistical analysis was performed using two-way ANOVA with Sidak’s mutiple comparison test. The percentage of viable cells is depicted as mean ± SEM. B The indicated cells were exposed to different concentrations of olaparib (0, 0.2, 1, 5, and 25 µM) for 120 h and MTT assay was performed. Whole-cell lysates were prepared and subjected to western blotting for the analysis of molecular response to olaparib. Trastuzumab-resistant (TR) cells are vulnerable to PARP inhibitionĪ Cells were treated with olaparib (0, 1, 5 µM) for 120 h.

F Representative images of human HER2-positive gastric cancer tissues stained with anti-PARP1 antibody (Left), and the corresponding H-score of the specimens (Right). E Cells were treated with trastuzumab (10 µg/ml) for the indicated period and subjected to immunoblotting. Data are shown as mean ± SEM, ns: not significant *p < 0.05 of three replicative experiments. Relative PARP1 expression was quantified and normalized to GAPDH expression by measuring the band intensity of PARP1 using ImageJ. D Cells were treated with MG132 (2 µM) for 12 h and lysed for western blotting. Three independent experiments were conducted, and the data are shown as mean ± SEM *, p < 0.05. The half-life of PARP1 was determined by measuring the band intensity of PARP1 using ImageJ. C Cells were treated with cycloheximide (50 µg/ml) for the indicated time and harvested for western blot of the chasing level of PARP1. Quantified PARP1 transcripts were normalized to β-actin, and the data of three biological replicative experiments are shown as mean ± SD, ns: not significant. B The mRNA expression level of PARP1 in TR cells and their counter parental cells was determined by reverse transcription-polymerase chain reaction (qRT-PCR). PARP1 is upregulated in trastuzumab-resistant (TR) cells, and functions distinctive from parental cellsĪ The expression level of PARP1, and pADPr in TR cells and their counter parental cells was determined by western blotting. Data of three independent experiments are presented as mean ± SD, ns: not significant, *p < 0.05 **p < 0.005. (Right) Each scattered dot represents the tail moment of single cell. (Left) Representative images of the assay are shown. C Cells treated with trastuzumab (10 µg/ml) for the indicated time period were subjected to comet assays. B Whole-cell lysate (WCL) was prepared and immunoblotted for determining the expression level of γ-H2AX. The percentage of γ-H2AX-positive cells was determined by examining n > 150 cells per sample. (Left) Representative images are presented, and the scale bar is 10 µm. After fixation, cells were permeabilized with 0.5% Triton X-100 and stained for γ-H2AX (red) and counterstained with DAPI (blue). However, sure usually stresses the subjective or intuitive feeling of assurance.TR cells sustains the functional DDR pathway to restore DSBs induced by trastuzumabĪ Cells were treated with trastuzumab (10 µg/ml) for the indicated periods. In some situations, the words sure and positive are roughly equivalent. You're always so cocksure about everything When could sure be used to replace positive? While in some cases nearly identical to positive, cocksure implies presumptuous or careless positiveness. Police are certain about the cause of the fire When can cocksure be used instead of positive? The meanings of certain and positive largely overlap however, certain may apply to a basing of a conclusion or conviction on definite grounds or indubitable evidence. I'm positive that's the person I saw When would certain be a good substitute for positive? While all these words mean 'having no doubt or uncertainty,' positive intensifies sureness or certainty and may imply opinionated conviction or forceful expression of it. Some common synonyms of positive are certain, cocksure, and sure. Frequently Asked Questions About positive How does the adjective positive contrast with its synonyms?

Positive syn