Journal: Frontiers in oncology

Article Title: Cordycepin Inhibits Triple-Negative Breast Cancer Cell Migration and Invasion by Regulating EMT-TFs SLUG, TWIST1, SNAIL1, and ZEB1.

doi: 10.3389/fonc.2022.898583

Figure Lengend Snippet: FIGURE 4 | Effects of cordycepin on the TWIST1 or SLUG reporter activity. (A) Luciferase reporter assay showed that CD decreased the TWIST1 reporter activity in Hela cells, as the relative light unit (RLU) was decreased with the increase in CD dosage. (B) Luciferase reporter assay showed that CD reduces the SLUG reporter activity in Hela cells, as the Relative Light Units (RLU) were reduced with the increase of CD dosage. *P < 0.05, **P < 0.01. (C, D) The levels of mRNA for TWIST1, SLUG, SNAIL1, and ZEB1 in 4T1 or BT549. 4T1 or BT549 cells were treated with 10 µM of CD for 24 h, total RNA was extracted, and quantitative RT-PCR analysis was performed. (E) The levels of mRNA for TWIST1, SLUG, SNAIL1, and ZEB1 in BT549 were treated with the indicated concentration of CD. BT549 cells were treated with 0, 5, 10, and 20 µM of CD for 24 h, total RNA was extracted, and semiquantitative RT-PCR and quantitative RT-PCR analyses were performed.

Article Snippet: SLUG (SC-166476), ZEB1 (SC-25388), and SNAI1 (SC-10432) antibodies were purchased from Santa Cruz (Dallas, TX, USA).

Techniques: Activity Assay, Luciferase, Reporter Assay, Quantitative RT-PCR, Concentration Assay, Reverse Transcription Polymerase Chain Reaction

Journal: Nature cell biology

Article Title: Snail/Slug-YAP/TAZ Complexes Control Skeletal Stem Cell Self-Renewal and Differentiation

doi: 10.1038/ncb3394

Figure Lengend Snippet: (a) Western blot of p-YAP, p-TAZ, p-Lats1 and p-Mst1 in SSCs isolated from Snail f/f /Slug +/+ and Snail f/f /Slug −/− mice and transduced with adeno-GFP or Cre. Results are representative of 3 experiments performed. (b) Association of YAP/TAZ with Snail was detected by immunoprecipitation in Cos-1 cells transfected with Flag-YAP, Flag-TAZ and Snail-HA, respectively, as indicated. Results are representative of 3 experiments performed. (c) Association of YAP/TAZ with Slug was detected by immunoprecipitation in Cos-1 cells transfected with Flag-YAP, Flag-TAZ and Slug-Myc, respectively, as indicated. Results are representative of 3 experiments performed. (d) Associations between endogenous YAP/TAZ and Snail/Slug were detected in SSCs cultured in the absence or presence of osteogenic medium. Results are representative of 3 experiments performed. (e) BMP2 increases Snail/Slug-YAP/TAZ complex formation. Calvarial osteoblast progenitors isolated from E17.5 mice were treated with 100 ng/ml BMP2 for 2 h. Endogenous Snail/Slug-YAP/TAZ complexes were detected by immunoprecipitation. Results are representative of 3 experiments performed. (f) In situ proximity ligation assay detection of endogenous Snail/YAP/TAZ and Slug/YAP/TAZ interactions in human SSCs. siRNAs: siGFP, siSNAIL, siSLUG or siTEADs(1–4) were transfected, respectively. Nuclei were counterstained with DAPI (blue). The detected interaction sites are marked by fluorescent dots (red). Scale bar: 5μm. Results are representative of 3 experiments performed.

Article Snippet: Human or mouse TEF-1, TEF-3, TEF-4 and TEF-5 (TEAD1–4) siRNAs, and human SNAIL or SLUG siRNAs were obtained from Santa Cruz Technology.

Techniques: Western Blot, Isolation, Transduction, Immunoprecipitation, Transfection, Cell Culture, In Situ, Proximity Ligation Assay