Journal: Nature Communications

Article Title: Transcriptional regulation of protein synthesis by mediator kinase represents a therapeutic vulnerability in MYC-driven medulloblastoma

doi: 10.1038/s41467-025-64937-3

Figure Lengend Snippet: a IC50 determination of various CDK8 inhibitors in MB cell lines. Unit: μmol. b IC50 of RVU120 at 72 h in MB and NHA cells. Experiments were performed in n = 3 independent experiments. All comparisons are to NHA cells. One-way ANOVA. c Dose-dependent proliferation curve of RVU120-treated primary MB cells from a G3-MB patient. n = 5 biological replicates. Mea n ± SEM. One-way ANOVA. d Immunofluorescence of CDK8 and DAPI. MB cells were treated with 1000 nM RVU120 for 48 h. Data points from n = 3 biological replicates. Scale bar, 10 μm. The line on the box plot represents the median. Two-sided unpaired t-test. e Immunoblot analysis of p-STAT1 following time-dependent treatment with 1000 nM RVU120 across MB cell lines. Representative of n = 3 experiments. f Immunoblot analysis of p-STAT1 levels following dose-dependent RVU120 treatment at 72 h. Representative of n = 3 experiments. Mea n ± SEM. Two-way ANOVA. g Methylcellulose assay in MB cells treated with RVU120. n = 3 biological replicates. Mea n ± SEM. Two-way ANOVA. h Annexin V apoptosis assay. MB cells were treated with 1000 nM RVU120 for 48 h. n = 3 biological replicates. Mea n ± SEM. Two-way ANOVA. i Identification of the brain tumor-initiating cell fraction in MB cells by ALDH expression demonstrates a decrease in the ALDH + fraction following 1000 nM RVU120 treatment for 48 h. n = 3 biological replicates. Mea n ± SEM. Two-way ANOVA. j Representative bioluminescence images of mice treated with RVU120 or vehicle (control, n = 8; RVU120, n = 6). k Kaplan–Meier survival curves of mice treated with vehicle ( n = 8) or RVU120 ( n = 6). Log-rank test. l Representative MRI of PDX411 xenograft mice treated with RVU120 or vehicle. Mice received the first scan after 14 days of treatment. Asterisks denote spongy tissue texture in mice. An adjusted texture analysis was performed to measure the tumor size. n = 3 mice. One control mouse died before the final scan. Mea n ± SEM. Two-way ANOVA.

Article Snippet: Antibodies used for western blot analysis were from the following sources: β-actin (Cell Signaling, 8457, 1:2000), CDK8 (Cell Signaling, 4101, 1:1000), 4EBP1 (Cell Signaling, 9644S, 1:1000), phospho-4EBP1 (Cell Signaling, 2855S, 1:1000), STAT1 (Cell Signaling, 9176S, 1:1000), phospho-STAT1 (Cell Signaling, 8826S, 1:1000), S6 (Cell Signaling, 2217 T, 1:1000), phospho-S6 (Cell Signaling, 4858 T, 1:1000), RNA Pol II (Cell Signaling, 2629S, 1:1000), and phospho-RNA Pol II-Ser2 (Cell Signaling, 13499, 1:1000).

Techniques: Immunofluorescence, Western Blot, Methylcellulose Assay, Apoptosis Assay, Expressing, Control

Journal: Nature Communications

Article Title: Transcriptional regulation of protein synthesis by mediator kinase represents a therapeutic vulnerability in MYC-driven medulloblastoma

doi: 10.1038/s41467-025-64937-3

Figure Lengend Snippet: a Dose-dependent assay of the combined treatment with RVU120 and Torin1 on day 5. Scale bar, 400 μm. n = 3 biological replicates . b Real-time proliferation assay quantifying the combined treatment with RVU120 and Torin1. Control ( n = 9); treatment ( n = 3) biological replicates. Mea n ± SEM. All p < 0.0001 vs. control (*). Two-way ANOVA. c Heatmap representation of the Fraction Affected and the Bliss interaction index across the five-point dose range of RVU120 and Torin1. Mean values of n = 3 biological experiments are shown. d The combination index of RVU120 and Torin1 using Chou-Talalay method. The mean combination index was determined from n = 3 biological replicates . e Apoptosis assay following combined treatment with RVU120 and Torin1. MB cells were treated for 48 h before staining with PI and Annexin V. n = 3 biological replicates. Mean ± SEM. Two-way ANOVA. f, g Effects of the combination of RVU120 and Torin1 on protein synthesis markers, phospho-Pol2 and phospho-STAT1, in MB cells after 48 h of treatment. Representative of n = 3 experiments. h The nude mice injected with D458 cells were treated with vehicle ( n = 10), RVU120 (40 mg/kg, n = 6), TAK-228 (1 mg/kg, n = 8), or their combination ( n = 7). i Representative Sagittal T2-weighted turboRARE MRI of D458 xenografted mice at 22 days. White arrows indicate tumors. MRI volumetric analysis is shown. n = 2 mice were selected and scanned in each group. j Kaplan–Meier survival analysis of mice treated with vehicle control ( n = 10), TAK-228 ( n = 8), RVU120 ( n = 6), or the combination of TAK-228 and RVU120 ( n = 7). Statistical significance was assessed using the log-rank test.

Article Snippet: Antibodies used for western blot analysis were from the following sources: β-actin (Cell Signaling, 8457, 1:2000), CDK8 (Cell Signaling, 4101, 1:1000), 4EBP1 (Cell Signaling, 9644S, 1:1000), phospho-4EBP1 (Cell Signaling, 2855S, 1:1000), STAT1 (Cell Signaling, 9176S, 1:1000), phospho-STAT1 (Cell Signaling, 8826S, 1:1000), S6 (Cell Signaling, 2217 T, 1:1000), phospho-S6 (Cell Signaling, 4858 T, 1:1000), RNA Pol II (Cell Signaling, 2629S, 1:1000), and phospho-RNA Pol II-Ser2 (Cell Signaling, 13499, 1:1000).

Techniques: Proliferation Assay, Control, Apoptosis Assay, Staining, Injection

Journal: Molecular Medicine Reports

Article Title: Regulatory role of POSTN in keloid pathogenesis

doi: 10.3892/mmr.2025.13701

Figure Lengend Snippet: Knockdown of POSTN blocks the JAK-STAT signaling pathway in KFs. (A) JAK-STAT pathway was revealed to be associated with POSTN knockdown in KFs by gene set enrichment analysis. (B) Heatmap showing DEGs related to the JAK-STAT pathway in si-POSTN and si-CTL KFs. Three independent RNA-sequencing experiments are shown in each group. (C) Reverse transcription-quantitative PCR analysis was performed to validate the DEGs related to the JAK-STAT pathway in si-POSTN- and si-CTL-transfected KFs (n=3). (D) Representative western blotting and semi-quantification of the protein levels of p-STAT1 (Tyr701) and p-STAT1 (Ser727) in KFs transfected with si-POSTN and si-CTL in the presence or absence of IFNα +β treatment (n=3). Data are presented as the mean ± SEM. ***P<0.001, **P<0.01, *P<0.05. DEGs, differentially expressed genes; KFs, keloid fibroblasts; NS, not significant; p-, phosphorylated; POSTN, periostin; si, small interfering.

Article Snippet: Antibodies against POSTN (cat. no. ab14041) and IL-4R (cat. no. ab203398) were from Abcam, antibodies against phosphorylated (p)-STAT1 (Tyr701) (cat. no. 9167), p-STAT1 (Ser727) (cat. no. 8826) and STAT1 (cat. no. 14994) were from Cell Signaling Technology, Inc., whereas the antibody against β-actin (cat. no. AF5003) was purchased from Beyotime Institute of Biotechnology.

Techniques: Knockdown, RNA Sequencing, Reverse Transcription, Real-time Polymerase Chain Reaction, Transfection, Western Blot