Journal: Oncotarget

Article Title: GDF15 contributes to radioresistance and cancer stemness of head and neck cancer by regulating cellular reactive oxygen species via a SMAD-associated signaling pathway

doi: 10.18632/oncotarget.13649

Figure Lengend Snippet: A, B. Administration of rhGDF15 increased the phosphorylated forms of SMAD family proteins in HNC cells. KB or FaDu cells were treated with serial doses of rhGDF15 (0-10 ng/ml) for 15 min (A) or 5 ng/ml rhGDF15 for various times (0-120 min) (n=3). (B) The cellular proteins were extracted and subjected to western blot analysis for SMAD family protein expressions (n=3). C. Effects of SMAD1-siRNA on the expressions of SMAD family proteins. After transfection of SMAD1-specific siRNA or the scramble oligonucleotides in HNC cells for 48h, cellular proteins were extracted for western blot analysis. GAPDH protein was used as an internal control (n=3). D. Silencing SMAD1 increased ROS level in HNC cells. KB or OECM1 cells were transfected SMAD1 specific siRNA or the scramble oligonucleotides for 48 h. After treating cells with 10 μM H 2 O 2 for 20 min, the Intracellular ROS levels were determined by DCF dye staining and analyzed with flow cytometry. E. Silencing SMAD1 suppressed spheroid cell formation in HNC cells. Fadu, OECM1 or KB cells were transfected SMAD1 specific siRNA or the scramble oligonucleotides for 48 h. These cells were then incubated in the spheroid cell culture condition and assessed after 14 days (n=3). F. The spheroid cell formation promoted by GDF15 was inhibited in SMAD knockdown HNC cells. KB or OECM1 cells were transfected SMAD1 specific siRNA or the scramble oligonucleotides for 48 h, with the addition of rhGDF15 (20 ng/ml). These cells were then incubated in the spheroid cell culture condition and assessed after 14 days (n=3). G. Effects of SMAD3-siRNA on the expressions of SMAD family proteins. After transfection of SMAD3-specific siRNA or the scramble oligonucleotides in HNC cells for 48h, cellular proteins were extracted for western blot analysis. GAPDH protein was used as an internal control (n=3). H. Silencing SMAD3 had no effect on ROS level in HNC cells. Fadu or OECM1 cells were transfected SMAD3-specific siRNA or the scramble oligonucleotides for 48 h. After treating cells with 10 μM H 2 O 2 for 20 min, the Intracellular ROS levels were determined by DCF dye staining and analyzed with flow cytometry. I. Silencing SMAD3 had no significant effect on spheroid cell formation in HNC cells. Fadu, OECM1 or Detroit cells were transfected SMAD3 specific siRNA or the scramble oligonucleotides for 48 h, with the addition of rhGDF15 (20 ng/ml). After 14 days of incubation in the spheroid cell culture condition, cells were assessed for spheroid formation (n=3). (*: p < 0.05, **: p < 0.01, ***: p < 0.001, n.s. : non-significance, t -test).

Article Snippet: The SMAD1-specific siRNA (#6223S) was purchased from Cell Signaling Technology (Danver, MA, USA).

Techniques: Western Blot, Transfection, Staining, Flow Cytometry, Incubation, Cell Culture

Journal:

Article Title: Chromosome Rearrangement and Diversification of Francisella tularensis Revealed by the Type B (OSU18) Genome Sequence

doi: 10.1128/JB.00506-06

Figure Lengend Snippet: Optical map comparisons of Francisella strains. EcoRI restriction maps for the type A Schu4 and ATCC 6223 and type B OSU18 chromosomes were constructed by OpGen (Madison, WI). Green indicates areas where the restriction maps are homologous for the paired genomes. Red in the ATCC 6223 genome indicates areas where the restriction maps have homologies in all three genomes. Red in the OSU18 and Schu4 genomes indicates regions in these genomes that are homologous to more than one region in ATCC 6223. The comparison of OSU18 to ATCC 6223 shows a level of rearrangement similar to that found when OSU18 was compared to Schu4 (Fig. ​(Fig.11 and ​and2).2). The comparison of Schu4 to ATCC 6223 demonstrates that rearrangement between chromosomes is prevalent within the type A subspecies, whereas the two finished type B genomes examined thus far (LVS and OSU18) suggest that rearrangements in this subspecies may be much less prevalent.

Article Snippet: Red in the ATCC 6223 genome indicates areas where the restriction maps have homologies in all three genomes.

Techniques: Construct