Journal: Cell Reports

Article Title: The Arabidopsis F-box protein FBW2 targets AGO1 for degradation to prevent spurious loading of illegitimate small RNA

doi: 10.1016/j.celrep.2022.110671

Figure Lengend Snippet:

Article Snippet: Anti cMyc-HRP , Miltenyi , Cat# 130-092-113, RRID: AB_871937.

Techniques: Virus, Recombinant, SYBR Green Assay, Western Blot, Protease Inhibitor, Isolation, Reverse Transcription, Magnetic Beads, Mass Spectrometry, Cloning, Sequencing, Software

Journal: Cancers

Article Title: A DNA Methylation-Dependent NOP56/MYC Positive Feedback Loop Promotes the Proliferation and Migration of Non-Small Cell Lung Cancer Through Regulating Ribosome Biogenesis

doi: 10.3390/cancers18050751

Figure Lengend Snippet: MYC mediates oncogenic role on NOP56 in NSCLC. ( A ) GSEA plot of MYC targets in NOP56-depleted A549 cells compared with control A549 cells. ( B ) qPCR analysis of MYC target gene expression in NOP56-depleted NSCLC cells or control. ( C ) Western blot analysis of MYC expression in NOP56-depleted NSCLC cells or control. ( D ) qPCR analysis of MYC expression in NOP56-depleted NSCLC cells or control. ( E ) Western blot analysis of MYC expression in MYC-depleted NSCLC cells or control. ( F – I ) CCK-8 assays, colony formation assays, transwell assays and wound healing assays in NSCLC cells with NOP56 overexpression or MYC knockdown. The data are shown as mean ± SD. Scale bar, 100 µM, * means ovNCS + siNC vs. ovNOP56 + siNC, * p < 0.05, ** p < 0.01, *** p < 0.001 by Student’s t test, # means ovNOP56 + siNC vs. ovNOP56 + siMYC, # p < 0.05, ## p < 0.01, by Student’s t test.

Article Snippet: Membranes were blocked with 5% nonfat milk and probed with primary antibodies at 4 °C overnight with primary antibodies against MYC (49 kDa, 1:2000, 10828-1-AP, Proteintech, Rosemont, IL, USA), NOP56 (66 kDa, 1:1000, A302-720A-T, Thermo, Waltham, MA, USA), or GAPDH (36 kDa, 1:5000, 60004-1-Ig, Proteintech, Rosemont, IL, USA).

Techniques: Control, Targeted Gene Expression, Western Blot, Expressing, CCK-8 Assay, Over Expression, Knockdown

Journal: Cancers

Article Title: A DNA Methylation-Dependent NOP56/MYC Positive Feedback Loop Promotes the Proliferation and Migration of Non-Small Cell Lung Cancer Through Regulating Ribosome Biogenesis

doi: 10.3390/cancers18050751

Figure Lengend Snippet: NOP56 regulates MYC through IRES-dependent translation. ( A ) RTL-P assays were conducted to detect the 2′-O methylation level of rRNA in NSCLC cells with NOP56 overexpression or control. ( B ) OP-Puro incorporation assay was conducted to detect the protein synthesis rate. ( C ) The bi-cistronic luciferase reporter was constructed as above and the Poliovirus (PV) IRES activity was calculated as the ratio of firefly luciferase activity over renilla luciferase activity. ( D ) The bi-cistronic luciferase reporter was constructed as above and the IRES-dependent translation (Fluc/Rluc) from IRES elements of MYC was measured. The data are shown as mean ± SD. Scale bar, 100 µM, * p < 0.05, ** p < 0.01 by Student’s t test.

Article Snippet: Membranes were blocked with 5% nonfat milk and probed with primary antibodies at 4 °C overnight with primary antibodies against MYC (49 kDa, 1:2000, 10828-1-AP, Proteintech, Rosemont, IL, USA), NOP56 (66 kDa, 1:1000, A302-720A-T, Thermo, Waltham, MA, USA), or GAPDH (36 kDa, 1:5000, 60004-1-Ig, Proteintech, Rosemont, IL, USA).

Techniques: Methylation, Over Expression, Control, Luciferase, Construct, Activity Assay

Journal: Cancers

Article Title: A DNA Methylation-Dependent NOP56/MYC Positive Feedback Loop Promotes the Proliferation and Migration of Non-Small Cell Lung Cancer Through Regulating Ribosome Biogenesis

doi: 10.3390/cancers18050751

Figure Lengend Snippet: MYC contributes to NOP56 transcriptional regulation. ( A – C ) Correlation between NOP56 mRNA and MYC mRNA expression in TCGA-LUSC, TCGA-LUAD and GSE30219 datasets. ( D ) The predicted MYC binding sites in the NOP56 promoter region. ( E ) qPCR analysis of NOP56 expression in MYC-depleted NSCLC cells or control. ( F ) The luciferase reporter was constructed as above and the transcription (Fluc) from WT/MUT promoter elements of MYC was measured. ( G ) ChIP assay was conducted to confirm the MYC binding sites in NOP56 promoter. The data are shown as mean ± SD. ** p < 0.01, *** p < 0.001 by Student’s t test.

Article Snippet: Membranes were blocked with 5% nonfat milk and probed with primary antibodies at 4 °C overnight with primary antibodies against MYC (49 kDa, 1:2000, 10828-1-AP, Proteintech, Rosemont, IL, USA), NOP56 (66 kDa, 1:1000, A302-720A-T, Thermo, Waltham, MA, USA), or GAPDH (36 kDa, 1:5000, 60004-1-Ig, Proteintech, Rosemont, IL, USA).

Techniques: Expressing, Binding Assay, Control, Luciferase, Construct