Journal: Blood Science
Article Title: DNAH2 facilitates the homologous recombination repair of Fanconi anemia pathway through modulating FANCD2 ubiquitination
Figure Lengend Snippet: The effect of DNAH2 knockdown on the viability of MMC-treated U2OS cells. (A) The U2OS cell line was transfected with the siDNAH2 or siFANCA, respectively. The siControl was utilized as the control. The RT-PCR assay was performed to measure the expression level of DNAH2 and FANCA , respectively. GAPDH gene was loaded as an internal reference gene. (B) The U2OS cell line was transfected with the siControl, siDNAH2, or siFANCA, respectively. Under the treatment of different concentrations of MMC (0, 50, 100, 150, 200, and 250 ng/mL), we measured the U2OS cell viability in the 3 groups, respectively. (C) and (D) In the presence (+) or absence (−) of 100 ng/mL MMC, we detected the status of the cell cycle and calculated the percentage of G2/M phase. The statistical difference between the MHC (−) and MHC (+) was analyzed by student t test. DNAH2 , Dynein Axonemal Heavy Chain 2; MMC: mitomycin C; RT-PCR: reverse transcription-polymerase chain reaction.
Article Snippet: 4.4 MMC sensitivity assay U2OS cell line was transfected with the siControl, siDNAH2, or siFANCA, respectively, and then treated by the different concentrations of MMC (0, 50, 100, 150, 200, 250 ng/mL in ddH2O, Sigma) for 24 h. After the adding of CellTiter 96 Aqueous One Solution Reagent (Promega) for 2 h, cellular viability was measured using an UltraMark Microplate Imaging System (Bio Rad, Hercules, CA) following the manufacturer's protocol.
Techniques: Transfection, Reverse Transcription Polymerase Chain Reaction, Expressing