alpha smooth muscle actin  (Cell Signaling Technology Inc)

Journal: Journal of Clinical and Translational Hepatology

Article Title: Single-cell RNA Sequencing Analysis Reveals That Targeting PLG–PLGRKT Signaling-mediated Pro-fibrotic Scar-associated Macrophages Ameliorates Liver Fibrosis in Biliary Atresia

doi: 10.14218/JCTH.2026.00278

Figure Lengend Snippet: (A) Representative immunohistochemical staining for CD9, TREM2, and PLGRKT in liver tissue sections from the study groups. (B) Multiplex immunofluorescence staining of PLGRKT, CD9, TREM2, α-SMA, and CD68 in liver tissues from BA and CC cohorts. DAPI was used for nuclear staining. α-SMA was used as a marker of activated fibroblasts, and CD68 was used as a macrophage marker. (C) Representative multiplex immunofluorescence images showing PLGRKT, α-SMA, and CD68+CD9+TREM2+ cells in liver tissue sections from different study groups. (D–E) Multiplex immunofluorescence staining of SAM-associated proteins (ANXA2, GPNMB, LGALS1, LGALS3, and SPP1) together with α-SMA in liver tissues from BA and CC cohorts. All images were acquired at ×40 magnification. Scale bars, 50 µm.α-SMA, alpha-smooth muscle actin; BA, biliary atresia; CC, choledochal cyst control; DC, disease control; DAPI, 4′,6-diamidino-2-phenylindole; SAMs, scar-associated macrophages; PLGRKT, plasminogen receptor with a C-terminal lysine.

Article Snippet: The following primary antibodies were used: alpha-smooth muscle actin (α-SMA; CST, 19245S; Cell Signaling Technology, Danvers, MA, USA; 1:600), CD68 (Abcam, ab213363; Cambridge, UK; 1:1,000), TREM2 (Abcam, ab223684; Cambridge, UK; 1:250), CD9 (Proteintech, 60232-1-g; Rosemont, IL, USA; 1:200), PLG-RKT (Thermo Fisher Scientific, PA5-98932; Waltham, MA, USA; 1:100), LGALS1 (Abcam, ab138513; Cambridge, UK; 1:10,000), SPP1 (Abcam, ab283656; Cambridge, UK; 1:2,000), LGALS3 (Abcam, ab76245; Cambridge, UK; 1:250), GPNMB (Abcam, ab222109; Cambridge, UK; 1:1,000), and ANXA2 (Abcam, ab178627; Cambridge, UK; 1:100)( ).

Techniques: Immunohistochemical staining, Staining, Tissue, Multiplex Assay, Immunofluorescence, Marker, Control

Journal: Signal Transduction and Targeted Therapy

Article Title: Breast cancer-associated fibroblast-derived exosomal miR-1290 promotes triple-negative breast cancer cell proliferation

doi: 10.1038/s41392-026-02922-y

Figure Lengend Snippet: miR-1290 is upregulated in breast cancer and BCAF/MB-231-derived exosomes. a Nanoparticle tracking analysis of exosomes isolated from BCAFs and MB-231 cells showing their size distribution. The mean size and standard deviation (SD) are indicated for each sample. b TEM images of exosomes isolated from BCAFs (upper) and MB-231 cells (lower) showing the characteristic morphology of the exosomes. Scale bar, 100 nm. c Western blot analysis of exosome markers (CD63, CD81, and flotillin-1), CAF markers (α-SMA and FAP) and cellular markers (GM130 and H3) in BCAF cells, MB-231 cells and their derived exosomes. d BCAF-derived exosome uptake by TNBC cells. PKH67-labeled BCAF-derived exosomes (green) were incubated with BT-549 and Hs578T cells for 1 hour. The cell nuclei were stained with Hoechst 33342 (blue). Scale bar, 20 μm. e Heatmap showing miRNA expression profiles in exosomes derived from BCAFs and MB-231 cells. Red indicates high expression, and blue indicates low expression. miR-1290 (highlighted in red) is highly expressed in both exosome types. f Volcano plots showing differentially expressed miRNAs between breast cancer or TNBC samples and normal controls in five independent GEO datasets ( GSE37963 , GSE45666 , GSE45498 , GSE38167 , and GSE61438 ). Red and blue dots represent upregulated and downregulated miRNAs, respectively, selected using fold-change and nominal p value cutoffs (|log2(fold change)| > 0.58 and nominal p < 0.05, two-tailed Welch’s t test). g Venn diagram analysis of upregulated miRNAs across the five GEO datasets revealed four commonly upregulated miRNAs. h Violin plot showing miR-1290 expression levels in serum samples from normal individuals ( N , n = 2686) and breast cancer patients ( P , n = 1280) from the GSE73002 dataset. The expression of miR-1290 was significantly upregulated in the breast cancer patient samples (*** p < 0.001, Mann‒Whitney U test). i Analysis of serum exosomal miR-1290 levels in normal ( N , n = 20) and TNBC patient ( P , n = 39) samples collected at Keimyung University Dongsan Hospital. The results support the potential of miR-1290 as a candidate diagnostic marker in an independent cohort (*** p < 0.001, Mann‒Whitney U test). j Kaplan‒Meier plot of overall survival for TNBC patients based on serum exosomal miR-1290 expression levels from Keimyung University Dongsan Hospital ( p = 0.006, log-rank test)

Article Snippet: Membranes were incubated with the following primary antibodies: CD63 (1:200; ABclonal, A5271), CD81 (1:200; ABclonal, A5270), flotillin-1 (1:1000; ABclonal, A6220), α-SMA (1:1000; ABclonal, A17910), FAP (1:1000; ABclonal, A23789 ), GM130 (1:1000; Abcam, ab52649), H3 (1:1000; Cell Signaling Technology, 4620S), ACTB (1:20,000; ABclonal, AC026), TBC1D4 (1:200; Cell Signaling Technology, 2670S), P70S6K (1:500; Cell Signaling Technology, 9202S), phospho-P70S6K (1:500; ABclonal, AP0564), and Ki67 (1:1000; Abcam, ab16667).

Techniques: Derivative Assay, Exosomes, Analysis, Isolation, Standard Deviation, Morphology, Western Blot, Labeling, Incubation, Staining, Expressing, Two Tailed Test, Blood/Plasma/Serum Samples, Diagnostic Assay, Marker

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Fisetin Attenuates Uterine Leiomyoma in Rats: Potential Crosstalk Between HMGB1, SphK1/S1P, and TGF‐β Signaling

doi: 10.1002/jbt.71105

Figure Lengend Snippet: Effect of fisetin on fibrosis biomarkers (TGF‐β1 and α‐SMA). (A) Immunostaining of TGF‐β1 and α‐SMA in the uterine tissues of different animal groups. In the control and FIS groups, TGF‐β1 expression was minimal in stromal cells and mononuclear immune cells. In the induction group, TGF‐β1 expression was prominent in stromal cells between muscle (green arrow) and within immune cells in the endometrial tissue (blue arrow). The UL + FIS group exhibited a significant decrease in TGF‐β1 expression in stromal and immune cells (blue arrow). Regarding α‐SMA, its expression in smooth muscle cells highlights myometrial thickness of approximately 4 mm and 4.5 mm in the control and FIS groups, respectively. In the induction group, α‐SMA expression indicates a notable increase in myometrial thickness, reaching 12 mm. In the UL + FIS group, α‐SMA expression shows a substantial reduction in myometrial thickness compared to the induction group, decreasing to about 6 mm (×40). (B) Relative mRNA expression of TGF‐β1. n = 10in each group. β‐actin was used to normalize expression data. Data are presented as means ± S.D. ** p ≤ 0.01, *** p ≤ 0.001 versus control group; $$ p ≤ 0.01, $$$ p ≤ 0.001 versus FIS‐treated group; ## p ≤ 0.01 versus UL group. FIS, fisetin; TGF‐β1, transforming growth factor β1; α‐SMA, α‐ α‐smooth muscle actin; UL, uterine leiomyoma; UL + FIS, uterine leiomyoma + fisetin.

Article Snippet: For immunohistochemistry analysis, formalin‐fixed, paraffin‐embedded tissue slices were stained for TGF‐β1 and α‐SMA antibodies supplied by Abclonal Inc., China (Cat. No: A16640 and A17910, for TGF‐β1 and α‐SMA, respectively) according to the manufacturer's instructions.

Techniques: Immunostaining, Control, Expressing