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: Journal of Translational Medicine

Article Title: Usp7 contributes to the tail regeneration of planarians via Islet/Wnt1 axis

doi: 10.1186/s12967-025-06134-w

Figure Lengend Snippet: Activation of Wnt signaling diminished the regeneration defect induced by Djusp7 inhibition. A The phenotype of the regenerative trunk fragments in planarian after both Usp7 RNAi and 10 nM Wnt agonist 1 treated (n = 10). Scale bar: 1 mm. B The mRNA expression level of marker genes of the regenerative trunk fragments in planarians at R 3 d. Vertical bars represent the mean ± SD (n = 3). C The expression of piwiA , prog1 , agat1 , and wnt1/P-1 in regenerative trunk worms by WISH (n = 6). Scale bar: 200 μm. D Immunostaining with anti-H3P antibody of the regenerative trunk fragments of the worms at R 3 d and R 7 d (n = 6). Scale bar: 200 μm. E Mitotic density in regenerative planarians. Vertical bars represent the mean ± SD (n = 6). F The protein expression level of USP7, H3P, Cyclin B1, BCL2, and Wnt1 in the trunk fragments after Wnt activation by Western blot. Asterisks indicate statistical differences (* p < 0.05; ** p < 0.01; *** p < 0.001)

Article Snippet: Wnt agonist 1 (Wnt signaling activator) (BML-284; 853,220–52-7) was purchased from Topscience.

Techniques: Activation Assay, Inhibition, Expressing, Marker, Immunostaining, Western Blot, Immunofluorescence, Real-time Polymerase Chain Reaction

Journal: Advanced Science

Article Title: Illuminating Mitochondrial RNA G‐Quadruplexes as Structural Brakes on RNA Granule Assembly and OXPHOS

doi: 10.1002/advs.202523462

Figure Lengend Snippet: A chemical genetic screen identifies the Wnt/β‐Catenin‐GRSF1‐mtRNA G4 axis regulating MRG assembly in cancer cells. (A) Depiction of the screening workflow. The data obtained regarding the fluorescence changes of MitoQUMA and MitoTO were standardized by calculating the Z‐ score for each inhibitor. (B) Z ‐score value for each inhibitor was plotted to identify hits with thresholds of | Z ‐score ( MitoQUMA ) |>3 and |Z‐score (MitoTO) | <0.5. (C) Live HeLa cells were treated with varying concentrations of ICG‐001, followed by staining with 2 µ m MitoQUMA . (D) Live HeLa cells were transfected with siRNA to knock down GRSF1 expression and then treated with or without 20 n m BML‐284, followed by staining with 2 µ m MitoQUMA . (E) Influence of MRG formation in HeLa cells upon siGRSF1 treatment with or without subsequent 20 nM BML‐284 exposure. MRGs were indicated by immunofluorescence staining of FASTKD2. For each sample of cell image, approximately 100 cells were measured. Biological replicates ( n = 3) were taken. The data are presented as mean ± SEM, and statistical significance is determined by the one‐way ANOVA with Dunnett's multiple comparisons test as (ns) not significant, ( * ) p < 0.05, ( ** ) p < 0.01, and ( *** ) p < 0.001. Scale bars for cell image: 10 µm.

Article Snippet: The treatment concentrations of IMT1 (TargetMol, T8841), Chloramphenicol (TargetMol, T1205), RHPS4 (TargetMol, T6967), Valinomycin (TargetMol, TP1072), Ionomycin (Beyotime, S1672), ICG‐001 (TargetMol, T6113) and BML‐284 (TargetMol, T8820) have been provided in the figure legend.

Techniques: Screening, Fluorescence, Staining, Transfection, Knockdown, Expressing, Immunofluorescence

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