Journal: eLife

Article Title: Uev1A counteracts oncogenic Ras stimuli in both polyploid and diploid cells

doi: 10.7554/eLife.107104

Figure Lengend Snippet: ( A ) Kaplan-Meier analysis of relapse-free survival in KRAS -mutant colorectal cancer patients with high or low expression levels of UBE2V1 and UBE2V2. ( B and E ) The knockdown efficiency assays. The relative mRNA levels were normalized to GAPDH . ( C–G ) Assays to evaluate the effects of UBE2V1- and UBE2V2-RNAi on colony formation and cell viability in SW480 and HCT116 cells. In ( B, C, E, and F ), three independent replicates were conducted, and statistical significance was determined using t test. In ( D and G ), five ( D ) and six ( G ) independent replicates were conducted at each time point, and statistical significance was determined using two-way ANOVA with multiple comparisons. * (p<0.05), *** (p<0.001), and **** (p<0.0001).

Article Snippet: The human colon cancer cell lines SW480 and HCT116, as well as the human embryonic kidney cell line 293T, were purchased from the American Type Culture Collection (ATCC), authenticated by STR profiling, and tested negative for mycoplasma contamination.

Techniques: Mutagenesis, Expressing, Knockdown

Journal: eLife

Article Title: Uev1A counteracts oncogenic Ras stimuli in both polyploid and diploid cells

doi: 10.7554/eLife.107104

Figure Lengend Snippet: ( A and C ) The knockdown efficiency assays. The relative mRNA levels were normalized to GAPDH . Three independent replicates were conducted, and statistical significance was determined using one-way ANOVA. ( B and D ) Assays to evaluate the effects of UBE2V1- and UBE2V2-RNAi on colony formation and cell viability in SW480 cells. Six independent replicates were conducted at each time point, and statistical significance was determined using two-way ANOVA with multiple comparisons. n.s . (p>0.05), * (p<0.05), ** (p<0.01), *** (p<0.001), and **** (p<0.0001).

Article Snippet: The human colon cancer cell lines SW480 and HCT116, as well as the human embryonic kidney cell line 293T, were purchased from the American Type Culture Collection (ATCC), authenticated by STR profiling, and tested negative for mycoplasma contamination.

Techniques: Knockdown

Journal: eLife

Article Title: Uev1A counteracts oncogenic Ras stimuli in both polyploid and diploid cells

doi: 10.7554/eLife.107104

Figure Lengend Snippet: ( A ) Western blotting to confirm the transient overexpression of UBE2V1 and UBE2V2 in SW480 and HCT116 cell lines. β-Actin was used as the loading control. ( B ) Western blotting to confirm the stable overexpression of UBE2V1 and UBE2V2 in SW480 cells, with UBE2V1-OE #1 and UBE2V2 #3 cell lines utilized in subcutaneous tumorigenesis assays. α-Tubulin was used as the loading control. In both ( A ) and ( B ), cells transfected with an empty overexpression vector served as the control. Figure 9—figure supplement 1—source data 1. PDF files that contain original western blots indicating the relevant bands and treatments. Figure 9—figure supplement 1—source data 2. Original files for western blot analysis.

Article Snippet: The human colon cancer cell lines SW480 and HCT116, as well as the human embryonic kidney cell line 293T, were purchased from the American Type Culture Collection (ATCC), authenticated by STR profiling, and tested negative for mycoplasma contamination.

Techniques: Western Blot, Over Expression, Control, Transfection, Plasmid Preparation

Journal: eLife

Article Title: Uev1A counteracts oncogenic Ras stimuli in both polyploid and diploid cells

doi: 10.7554/eLife.107104

Figure Lengend Snippet: ( A and B ) 5-Ethynyl-2’-deoxyuridine (EdU) incorporation assays to assess the effects of UBE2V1 and UBE2V2 overexpression (OE) on cell proliferation in SW480 and HCT116 cells. Empty OE vector was used as the control. All images in ( A ) are of the same magnification. ( C–E ) Assays to evaluate the effects of UBE2V1- and UBE2V2-OE on colony formation and cell viability in SW480 and HCT116 cells. In ( B and D ), three independent replicates were conducted, and statistical significance was determined using one-way ANOVA. In ( E ), five independent replicates were conducted, and statistical significance was determined using two-way ANOVA with multiple comparisons. * (p<0.05), ** (p<0.01), and **** (p<0.0001).

Article Snippet: The human colon cancer cell lines SW480 and HCT116, as well as the human embryonic kidney cell line 293T, were purchased from the American Type Culture Collection (ATCC), authenticated by STR profiling, and tested negative for mycoplasma contamination.

Techniques: Over Expression, Plasmid Preparation, Control