Journal: Journal of Virology

Article Title: The Epstein-Barr Virus (EBV)-Induced Tumor Suppressor MicroRNA MiR-34a Is Growth Promoting in EBV-Infected B Cells

doi: 10.1128/JVI.07056-11

Figure Lengend Snippet: Canonical miR-34a growth control targets are poorly expressed in LCLs relative to HCT-116 cells. (a) Relative mRNA levels as determined by qRT-PCR of cyclin D1, D2, and E2; cdk4; cdk6; and c-Met in LCLs and HCT-116 cells. The asterisk indicates a value below the threshold for detection. (b) Western analysis of miR-34a targets in HCT-116 cells and EF3D LCLs expressing pcDNA3 vector (−) or pCDNA3-miR-34a (+). Quantitation of protein expression is noted below each band.

Article Snippet: The antibodies used were cyclin D1 and D2 (BD; 554203), cyclin E2 (Cell Signaling; 4132), cdk4 (Santa Cruz; H-22), cdk6 (Santa Cruz; C-21), and Met (Cell Signaling; 4560).

Techniques: Control, Quantitative RT-PCR, Western Blot, Expressing, Plasmid Preparation, Quantitation Assay

Journal: bioRxiv

Article Title: Proteomic Analysis Reveals Trilaciclib-Induced Senescence

doi: 10.1101/2024.03.12.584620

Figure Lengend Snippet: A) The fitness effects of CRISPR-Cas9 gene knockout on TP53 , CDK4 , CDK6 , and RB1 genes (DepMap Public+Score, Chronos, https://depmap.org/portal/ )) were correlated with trilaciclib sensitivity across 694 cell lines. Pearson correlation coefficient (r) and p-value are shown in each graph. B-C) The sensitivity of trilaciclib varies across different histologic cancer types (DepMap, BRD: BRD-K00003412-300-01-9, PRISM Repurposing Public 23Q2; https://depmap.org/portal/ ). In C) blood cancers are subdivided into Hodgkin lymphoma (HL, 2 cell lines), acute myeloid leukaemia (AML, 21 cell lines), T-cell acute lymphoblastic leukaemia/ lymphoma (T-ALL/LBL, 10 cell lines), myeloproliferative neoplasms (MPN, 12 cell lines), non-Hodgkin lymphoma (NHL, 54 cell lines), and B-cell acute lymphoblastic leukaemia/ lymphoma (B-ALL/LBL, 9 cell lines). D) The inhibition concentration at 50 % cell death (IC50) curves were analysed in K562, JURKAT, U937, MOLT-4, and NCI-H929 cells after 72 hours of treatment. Standard deviation of four biological replicates is shown. E) Percentage of live cells (Annexin V negative and 7-AAD negative cells, light blue), apoptotic cells (Annexin V positive and 7-AAD negative cells, purple), and necrotic cells (7-AAD positive cells, pink) after 72 hours of treatment. The statistical significance of the comparisons with resting is indicated as follows: ‡, P ≤ 0.001; ns, not significant. Standard deviation of three biological replicates is shown.

Article Snippet: Then, it was probed with phospho-Rb (Ser 807/811) (#8516, Cell Signaling), p21 (#2947, Cell Signaling), CDK4 (sc-23896, Santa Cruz), CDK6 (#13331, Cell Signaling), α-Tubulin (T9026) (Sigma-Aldrich) antibodies overnight at 4 °C.

Techniques: CRISPR, Gene Knockout, Inhibition, Concentration Assay, Standard Deviation

Journal: Cancer cell international

Article Title: MicroRNA-550a-3-5p controls the brain metastasis of lung cancer by directly targeting YAP1.

doi: 10.1186/s12935-021-02197-z

Figure Lengend Snippet: Fig. 7 The effects of miR-550a-3-5p on cleaved-PARP, pRB, CDK6, YAP1, CTGF, and CYR61. A Western blotting images. B Cleaved-PARP protein expression. C pRB protein expression. D CDK6 protein expression. E YAP1 protein expression. F CTGF protein expression. (G) CYR61 protein expression. *P < 0.05, compared with the control group; #P < 0.05, compared with the miR-550a-3-5p mimics group

Article Snippet: After blocking with 5% skim milk, the membranes were incubated with anti-cleaved-Poly(ADP-ribose) polymerase (PARP) antibody (1:1000, Abcam), anti-RB transcriptional corepressor 1 (pRB) antibody (1:1000, Abcam), anti-cyclin dependent kinase 6 (CDK6) antibody (1:1000, Abcam), anti-Yes1 associated transcriptional regulator (YAP1) antibody (1:1000, Abcam), anti- connective tissue growth factor (CTGF) antibody (1:1000, Proteintech Group, Inc.), anti-cysteine rich angiogenic inducer 61 (CYR61) antibody (1:1000, Abcam), and anti-GAPDH antibody (1:10,000, Proteintech Group, Inc.) overnight at 4 °C.

Techniques: Western Blot, Expressing, Control

Journal: Oncology reports

Article Title: miR-3619-5p inhibits prostate cancer cell growth by activating CDKN1A expression.

doi: 10.3892/or.2016.5250

Figure Lengend Snippet: Figure 6. miR-3619-5p inhibits cell cycle related gene expression primarily by regulating CDKN1A expression. DU145 and PC3 cells were transfected with 50 nM of the indicated RNAs for 72 h. (A) Expression of p21 mRNA was assessed by real-time PCR. GAPDH served as a loading control. (B) Induction of p21 protein expression was detected by western blot analysis. GAPDH were also detected and served as a loading control. (C) Expression of cyclin D1, CDK4 and CDK6 mRNA was detected by real-time PCR. GAPDH served as a loading control. (D) Expression of cyclin D1, CDK4 and CDK6 mRNA was determined by western blot analysis. α-Tublin served as a loading control. *P<0.05, **P<0.01 and ***P<0.001.

Article Snippet: After blocking the membranes were incubated overnight at 4 ̊C with appropriate dilutions of specific primary antibodies as follows: p21 (1/2000) (Cell Signaling Technology), cyclin D1 (1/2000) (Affinity, USA), CDK4 (1/1000) (Affinity), CDK6 (1/2000) (Affinity), GAPDH (1/500) (Boster, Wuhan, China) and α-tubulin (1/500) (Boster).

Techniques: Gene Expression, Expressing, Transfection, Real-time Polymerase Chain Reaction, Control, Western Blot