complementary dna microarray  (Agilent technologies)

Journal: Cancer Science

Article Title: Induction of DNA strand breaks is critical to predict the cytotoxicity of gemtuzumab ozogamicin against leukemic cells

doi: 10.1111/j.1349-7006.2012.02343.x

Figure Lengend Snippet: Protein expression levels of DNA repair proteins. Genes found to be altered in two gemtuzumab ozogamicin (GO)‐resistant subclones (HL/GO, HL/GO‐CSA) were evaluated for their protein expression levels by Western blotting.

Article Snippet: The hybridization was carried out between Cy‐3‐labeled total RNA from the parental HL‐60 cells and Cy‐5‐labeled total RNA from the HL/GO cells or HL/GO‐CSA cells on microarrays of complementary DNA that contained 35 000 elements (Operon Aros, Human Genome Oligo Set, Version 4.0; Operon Biotechnologies, Tokyo, Japan).

Techniques: Expressing, Western Blot

Journal: Cancer Science

Article Title: Induction of DNA strand breaks is critical to predict the cytotoxicity of gemtuzumab ozogamicin against leukemic cells

doi: 10.1111/j.1349-7006.2012.02343.x

Figure Lengend Snippet: Relationship between gemtuzumab ozogamicin (GO)‐associated parameters and GO sensitivity. (A) Tail moment values (number of DNA strand breaks) and IC 50 values were plotted within the same cell line (HL‐60, HL/GO, HL/GO‐CSA, HL/Ara‐CDNR, K562). The IC 50 values were determined using the XTT assay. Tail moment values were determined by the Comet assay after cells had been treated with 10 μg/mL GO for 6 h. The values are the means of triplicate determinations. (B–D) Similarly, the correlation between the IC 50 values and each of the factors (CD33 positivity, P‐glycoprotein [P‐gp] expression, and multridrug resistance protein‐1 [MRP1] mRNA) were evaluated. (E) Comparison was made between the CD33‐positive/P‐gp‐negative/MRP1‐negative cell lines (HL‐60, K562) and the others (HL/GO, HL/GO‐CSA, HL/ara‐CDNR). Bars represent the means.

Article Snippet: The hybridization was carried out between Cy‐3‐labeled total RNA from the parental HL‐60 cells and Cy‐5‐labeled total RNA from the HL/GO cells or HL/GO‐CSA cells on microarrays of complementary DNA that contained 35 000 elements (Operon Aros, Human Genome Oligo Set, Version 4.0; Operon Biotechnologies, Tokyo, Japan).

Techniques: XTT Assay, Single Cell Gel Electrophoresis, Expressing, Comparison