Journal: Military Medical Research

Article Title: Glucocorticoids trigger muscle-liver crosstalk to attenuate acute liver injury and promote liver regeneration via the FGF6-FGFBP1 axis

doi: 10.1186/s40779-025-00618-y

Figure Lengend Snippet: FGF5 and FGF5s compete for binding to FGFBP1 via LLPS and regulate liver regeneration. a BrdU cell proliferation assay results to detect cell proliferation ability in AML12 cells overexpressing the 18 FGFs and treated with rFGFBP1 (10 ng/ml) or rFGFBP1 and FGFBP1Ab (2 μg/ml) ( n = 3). b Hepatic protein levels of FGF5 and FGF5s at the indicated times after partial (2/3) hepatectomy (PHx), with the corresponding quantification ( n = 2). c Representative liver FGFBP1 and FGF5 immunofluorescence results 24 h after acetaminophen (APAP) or saline administration. d Domain structure and the intrinsically disordered tendency of mouse FGF5 and FGF5s. IUPred assigned scores of the disordered tendencies of the sequences between 0 and 1, with scores higher than 0.5 indicating disorder. The amino acid sequence of FGF5 contains one IDR. e Confocal microscopy images of the assembly status of 5 μmol/L purified rFGF5-mCherry, rFGF5s-mCherry, or rΔIDR-mCherry with 0 or 10% PEG. Scale bar = 5 µm. f Images of purified protein colocalization assay on the cell surface via confocal microscopy and representative curves describing the distribution of the relative fluorescence intensities of FGFBP1 (green) and FGF5 or FGF5s (red). Top two panels: rFGFBP1-EGFP (5 nmol/L) with rFGF5-mCherry (20 nmol/L) or rFGF5s-mCherry (10 nmol/L). Bottom three panels: rFGFBP1-EGFP (5 nmol/L) with rFGF5-mCherry (20 nmol/L) and different amounts of rFGF5s (10, 20, or 40 nmol/L). Scale bar = 5 µm. g Representative albumin (ALB, a hepatocellular function marker) immunofluorescence of human liver organoids (HLOs) in the indicated groups. h Measurement of ALB secretion in HLOs ( n = 3). i Size calculation of HLOs in the indicated groups (CON, n = 146; rFGFBP1, n = 157; rFGFBP1 + rFGF5, n = 184). * P < 0.05, *** P < 0.001. FGF5 fibroblast growth factor 5, FGF5s short form of FGF5, FGFBP1 fibroblast growth factor binding protein 1, LLPS liquid–liquid phase separation, IDR intrinsically disordered region, PEG polyethylene glycol, EGFP enhanced green fluorescent protein, DIC differential interference contrast, DAPI 4’,6-diamidino-2-phenylindole, SP signal peptide

Article Snippet: Cell proliferation was detected using a BrdU Cell Proliferation Assay Kit (6813S, Cell Signaling Technology, MA, USA) according to the manufacturer’s instructions.

Techniques: Binding Assay, BrdU Cell Proliferation Assay, Immunofluorescence, Saline, Sequencing, Confocal Microscopy, Purification, Fluorescence, Marker

Journal: Frontiers in Cardiovascular Medicine

Article Title: TNIK-driven regulation of ERK5 transcriptional activity in endothelial cells

doi: 10.3389/fcvm.2025.1526676

Figure Lengend Snippet: TNIK promotes EC proliferation through a MEK5-dependent mechanism: BrdU incorporation assays were performed in HUVECs to evaluate the effects of TNIK WT and DN-MEK5 on cell proliferation. Overexpression of TNIK WT or DN-MEK5 individually increased BrdU incorporation relative to vector control. Co-transfection of TNIK WT and DN-MEK5 attenuated this proliferative effect, indicating that intact MEK5 signaling is required for TNIK-mediated promotion of EC proliferation. Data are presented as mean ± SEM from four independent experiments. Statistical comparisons were performed using one-way ANOVA followed by post hoc testing; Significance thresholds ** p < 0.01; * p < 0.05.

Article Snippet: To assess the effect of TNIK-ERK5 signaling on EC proliferation, a bromodeoxyuridine (BrdU) incorporation assay was performed using a commercial BrdU Cell Proliferation Assay Kit (Cell Biolabs, #CBA-251).

Techniques: BrdU Incorporation Assay, Over Expression, Plasmid Preparation, Control, Cotransfection

Journal: Cell Communication and Signaling : CCS

Article Title: C/EBPβ increases tumor aggressiveness by enhancing KIFC1 expression in androgen receptor negative triple negative breast cancer

doi: 10.1186/s12964-025-02243-7

Figure Lengend Snippet: Silencing of C/EBPβ and KIFC1 reduces proliferation in AR-TNBC cells. ( A – G ) Representative immunofluorescence images ( A – D ) and quantification bar graphs ( E – G ) showing BrdU (green) incorporation in AR + TNBC ( A , B ) and AR-TNBC ( C , D ) cells transfected with scrambled or C/EBPβ siRNA ( A – D , E ) or treated with CW069 ( A – D , G ). Nuclei were counterstained with Hoechst (blue) and α-tubulin (red). ( H – J ) Bar graphs showing BrdU incorporation in cells transfected with scrambled siRNA, C/EBP siRNA ( H ), KIFC1 siRNA ( I ), or treated with CW069 ( J ). Absorbance was measured at 450–540 nm. A : HCC70, B : MFM223, C : HCC1806, D : BT20; A , B -AR-positive TNBC, C , D-AR-negative TNBC. Bars indicate mean ± SEM. Unpaired two-tailed Student’s t -test with Welch’s correction was used to determine statistical significance. * P < 0.05, ** P < 0.005, ns = non-significant

Article Snippet: BrdU cell proliferation kit , 2750 , EMD Millipore.

Techniques: Immunofluorescence, Transfection, BrdU Incorporation Assay, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: CBD promotes antitumor activity by modulating tumor immune microenvironment in HPV associated head and neck squamous cell carcinoma

doi: 10.3389/fimmu.2025.1528520

Figure Lengend Snippet: CBD treatment promotes anti-cancer activity in HPV-POSITIVE HNSCC cells. Measurement of cell proliferation by AquaBluer Assay after treatment with 10 μM of CBD in HPV-positive HNSCC cells (A) UPCI: SCC154 and (B) UD-SCC-2 for 24 and 48 hours. Graphical representations of percentage of cell proliferation by BrdU assay on (C) UPCI: SCC154 and (D) UD-SCC-2 after treatment with CBD 10 μM for a time period of 24 h and 48h. Cell proliferation decreases significantly with CBD treatment compared to Vehicle treated group. Measurement of percentage of apoptosis post CBD treatment for 48 hours in HPV-positive HNSCC cells (E) UPCI: SCC154 and (F) UD-SCC-2. Measurement of percentage of cellular migration post CBD treatment for 48 hours in HPV-positive cells (G) UPCI: SCC154 and (H) UD-SCC-2. Statistical analysis was performed by unpaired Student’s t-test. [*p<0.05 **p<0.01].

Article Snippet: After incubation with drugs, the cells were incubated with 1X BrdU for 24 h. The BrdU assay kit was purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Activity Assay, BrdU Staining, Migration