Journal: Cancer Cell International

Article Title: Sestrin2 reduces cancer stemness via Wnt/β-catenin signaling in colorectal cancer

doi: 10.1186/s12935-022-02498-x

Figure Lengend Snippet: The effects of Sestrin2 on cancer stemness are mediated by the Wnt/β-catenin pathway. A Western blot analysis of Wnt/β-catenin signaling-related proteins. The relative protein expression is shown in the right panel. β-Actin was used as a loading control * P = 0.05; Mann–Whitney test; lines showed medians). B Sphersphere formation assay in LV-Sestrin2 and LV-GFP CRC cells with or without exposure to BML-284 (0.1 µM). C Number of spheres, * compared with LV-GFP, * P = 0.05; # compared with LV-Sestrin2, # P = 0.05; Mann–Whitney test; lines showed medians. D Western blot analysis of proteins related to the Wnt/β-catenin pathway (β-catenin, c-Myc) and cancer stemness (CD44) in LV-Sestrin2 and LV-GFP CRC cells with or without incubation with BML-284 (0.5 µM) for 24 h in HCT-116 cells. E The relative protein expression. * Compared with LV-GFP, * P = 0.05, # compared with LV-Sestrin2, # P = 0.05, Mann–Whitney test; lines showed medians

Article Snippet: HCT-116 cells in the LV-Sestrin2 and LV-GFP groups were treated with 0.5 μM BML-284 (Wnt signaling activator; MedChemExpress, Monmouth Junction, NJ, USA) in dimethyl sulfoxide for 24 h and then collected for western blot analysis.

Techniques: Western Blot, Expressing, Control, MANN-WHITNEY, Tube Formation Assay, Incubation

Journal: bioRxiv

Article Title: Metavinculin mediates differentiation cues into myogenic gene expression

doi: 10.1101/2024.02.21.578728

Figure Lengend Snippet: a Relative expression of canonical Wnt target genes Myc, Ccnd1, Ascl2, Id2, Tcf4 and Mmp9 in C2C12 cells cultured under various conditions. b Western blot showing canonical Wnt mediator Ctnnb (β-catenin) protein level in C2-Scr and C2-shmVcl2 cells cultured as indicated. c Co-immunoprecipitation analysis showing direct vinculin and Ctnnb interaction in C2-Scr and C2-shmVcl2 cells grown under indicated conditions. d Bar graph showing RT-qPCR validation of Wnt target genes activation by canonical Wnt activator BML-284 in C2C12 cells. e Relative expression of myogenic differentiation associated genes in C2C12 cells treated with BML-284 or vehicle in DM media shown in RT-qPCR. f Myogenic protein expression in BML-284 or vehicle treated C2C12 cells cultured under DM conditions is shown in western blot. g Representative images of Myh stain (green) and DAPI (blue) in C2C12 cells treated with BML-284 or vehicle, Scale bar 50 μM. Fusion index is shown in a bar graph. Actb act as a loading control in western blot. Data are shown as mean value ±SEM, n=3. p value * ≤0.05; **≤0.01; ***≤0.001; ****≤0.0001.

Article Snippet: Wnt activator BML-284 (5 mM) (AdooQ, USA) or inhibitor KY02111 (10 mM) (AdooQ, USA) and vehicle was treated in indicated culturing conditions and refreshed during media change.

Techniques: Expressing, Cell Culture, Western Blot, Immunoprecipitation, Quantitative RT-PCR, Activation Assay, Staining