Journal: Developmental cell

Article Title: The UBE2F-CRL5 ASB11 -DIRAS2 axis is an oncogene and tumor suppressor cascade in pancreatic cancer cells.

doi: 10.1016/j.devcel.2024.03.018

Figure Lengend Snippet: Figure 1. High UBE2F expression correlates with poor survival of PDAC patients and UBE2F regulates growth and survival of pancreatic cancer cells (A and B) UBE2F mRNA is overexpression in PDAC tissues, which correlates with poor patient survival. UBE2F mRNA levels in normal and PDAC tissues were analyzed in GEPIA database (A). The PDAC tissue microarrays were stained for UBE2F levels, and the survival of PDAC patients with high and low UBE2F expression were analyzed. The Kaplan-Meier survival analysis of PDAC patients with high and low UBE2F expression were shown. **p < 0.01 (B). (C–E) UBE2F knockdown inhibits the growth and survival of pancreatic cancer cells. PANC1 cells were transfected with indicated siRNAs and then subjected to IB (C), Cell counting kit-8 (CCK-8) (D), and clonogenic survival assays (E). (F–H) UBE2F overexpression promotes the growth and survival of pancreatic cancer cells. PANC1 cells were transfected with indicated constructs and then subjected to IB (F), CCK-8 (G), and clonogenic survival assays (H). HA-UBE2F-MUT: HA-UBE2F- C116A. Data shown are mean ± SEM of three in- dependent experiments. *p < 0.05, **p < 0.01, two- sided, unpaired Student’s t test. See also Figure S1.

Article Snippet: Cell proliferation was measured using Cell Counting Kit-8 (CCK-8, Cat No. HY-K0301, MedChem Express) or 1-step luminescence ATP detection assay system (Perkin Elmer) according to the manufacturer’s instructions.

Techniques: Expressing, Over Expression, Staining, Knockdown, Transfection, Cell Counting, CCK-8 Assay, Construct

Journal: Developmental cell

Article Title: The UBE2F-CRL5 ASB11 -DIRAS2 axis is an oncogene and tumor suppressor cascade in pancreatic cancer cells.

doi: 10.1016/j.devcel.2024.03.018

Figure Lengend Snippet: Figure 6. DIRAS2 mediates UBE2F regulation of growth of pancreatic cancer cells (A–F) DIRAS2 is a mediator of UBE2F growth regulation in pancreatic cancer cells harboring a mutant RAS. PANC1 and Miapaca-2 cells were transfected with indicated siRNA oligos for 48 h. Cells were then subjected to IB (A and B), CCK-8 (C and D), and colony formation (E and F) assays. Data shown are mean ± SEM of three independent experiments. *p < 0.05, **p < 0.01. (G–L) The UBE2F-DIRAS2 axis has no effect on pancreatic cancer cells harboring WT RAS. BxPC3 cells were transfected with indicated siRNA oligos for 48 h. Cells were then subjected to IB (G and J), CCK-8 (H and K), and colony formation (I and L) assays. Ns, no significance. See also Figures S5 and S6.

Article Snippet: Cell proliferation was measured using Cell Counting Kit-8 (CCK-8, Cat No. HY-K0301, MedChem Express) or 1-step luminescence ATP detection assay system (Perkin Elmer) according to the manufacturer’s instructions.

Techniques: Mutagenesis, Transfection, CCK-8 Assay

Journal: Viruses

Article Title: Stromal Antigen 2 Deficiency Induces Interferon Responses and Restricts Porcine Deltacoronavirus Infection

doi: 10.3390/v14081783

Figure Lengend Snippet: Establishment of STAG2-depleted IPEC-J2 cells line. ( A ) Western blot analysis for STAG2 expression in IPEC-J2 cell line; ( B ) Gene sequence alignment revealed that were heterozygous for STAG2 knockout alleles generated using CRISPR-Cas9 gene editing; ( C ) Cell viability was determined using CCK-8 detection; ( D ) Western blot analysis for STAG2 expression of WT IPEC-J2, STAG2 − / − IPEC-J2 passage 5 (P. 5), passage 10 (P. 10), passage 15 (P. 15) and passage 20 (P. 20).

Article Snippet: Cell viabilities were assessed using a cell counting kit-8 (CCK-8) (Cat NO. GK10001, GLPBIO, Montclair, CA, USA).

Techniques: Western Blot, Expressing, Sequencing, Knock-Out, Generated, CRISPR, CCK-8 Assay