Journal: International Journal of Oral Science

Article Title: Nociceptive adenosine A 2A receptor on trigeminal nerves orchestrates CGRP release to regulate the progression of oral squamous cell carcinoma

doi: 10.1038/s41368-024-00308-w

Figure Lengend Snippet: Presence of nociceptive nerves in adenosine-concentrated OSCC. a The upper panel presents the broad view of immunostaining of TRPV1 in the patient cohort collected by West China Hospital of Stomatology (WCHS). Scale bar: 2 mm. The lower panel denotes representative samples with and without TRPV1 neuronal infiltration (‘TRPV1 positive’ and ‘TRPV1 negative’). Scale bar: 50 µm. b Kaplan–Meier plot delineating survival probability for WCHS-OSCC patients stratified against TRPV1 neuron infiltration in their tumors, n = 111 patients. c Comparison of adenosine concentration in normal epithelium and HSC3 xenograft of mice, represented as net relative fluorescence unit (RFU). n = 3 mice. d Immunostaining of CD73 in human normal epithelium and human OSCC cells. Scale bars, 50 μm. e Kaplan–Meier plot delineating survival probability for TCGA-HNSC patients stratified against NT5E expression in their tumors. NT5E -high and NT5E -low groups are defined as above or below the median of NT5E expression. The TCGA-HNSC cohort database analyzed was updated to March 29th, 2023, n = 518 patients. Statistical analysis was conducted using unpaired Student’s t -test ( c ) and Log-rank test ( b , e )

Article Snippet: For non-fluorescent staining, antigen was retrieved either through 1X Tris-EDTA (for CD73, p-ERK and YAP) or 10 mM citric acid (for TRPV1and Ki67) before incubated with corresponding primary antibody TRPV1 (Alomone labs ACC-030), CD73(CST 13160 S), p-ERK (CST 4370 S), Ki67 (HUABIO HA721115) and YAP (CST 14074) in blocking solution, followed by reaction enhancer solution and horseradish peroxidase (HRP)-conjugated secondary antibody (ORIGENE PV-9001).

Techniques: Immunostaining, Comparison, Concentration Assay, Fluorescence, Expressing

Journal: NPJ Regenerative Medicine

Article Title: CD73 overexpression in ADSCs accelerates bladder repair by regulating the NFκB/NLRP3/caspase-1 signaling axis in neurogenic bladder rats

doi: 10.1038/s41536-026-00454-1

Figure Lengend Snippet: A Flow cytometry results of CD73-positive ADSCs. B Flow cytometry results of CD73-negative ADSCs. C In vitro, immunofluorescence staining of ADSCs in each group. Red indicates CD73, green indicates VEGF, blue indicates DAPI, scale bar = 20 µm. D Quantitative analysis of immunofluorescence staining results among different groups ( n = 3). E Western blot results of CD73 and VEGF proteins after transfection or APCP treatment in each group. F Quantitative analysis of Western blot results. G CCK-8 assay results among different groups of ADSCs in conditioned medium ( n = 3). H Colony formation assay results among RBSMC in conditioned medium ( n = 3). I Cell migration assay results and quantitative analysis among different groups of ADSCs in conditioned medium ( n = 3). J Wound healing assay results among RBSMC in conditioned medium ( n = 3). Data presented as ±SD. * and # indicates P < 0.05, ** and ## indicates P < 0.01, and *** and ### indicates P < 0.001. Full-length blots/gels are presented in Supplementary Fig. .

Article Snippet: The samples were subjected to SDS-PAGE, and primary antibodies were used against CD73 (diluted 1:1000, Affinity, Ontario, Canada), VEGF (diluted 1:1000, proteintech, Chicago, IL, USA), SDF-1 (diluted 1:1000, proteintech, Chicago, IL, USA), NLRP3 (diluted 1:1000, Affinity, Ontario, Canada), p-AKT (diluted 1:1000, Affinity, Ontario, Canada), AKT (diluted 1:1000, Affinity, Ontario, Canada),caspase-1 (diluted 1:1000, Servicebio, Wuhan, Hubei, China), p-NFκB (diluted 1:1000, Cell Signaling Technology, Danvers, MA, USA), NF-κB (diluted 1:1000, Cell Signaling Technology, Danvers, MA, USA), and GAPDH (diluted 1:1000, Abcam, Cambridge, UK).

Techniques: Flow Cytometry, In Vitro, Immunofluorescence, Staining, Western Blot, Transfection, CCK-8 Assay, Colony Assay, Cell Migration Assay, Wound Healing Assay

Journal: NPJ Regenerative Medicine

Article Title: CD73 overexpression in ADSCs accelerates bladder repair by regulating the NFκB/NLRP3/caspase-1 signaling axis in neurogenic bladder rats

doi: 10.1038/s41536-026-00454-1

Figure Lengend Snippet: A Western blot results of CD73 and VEGF in the bladder tissues of NB + CD73⁺ / ⁺ group on days 0, 7, 14, 21, and 28 after treatment. B Quantitative analysis of the Western blot results for CD73 and VEGF ( n = 3). C The results of the mean pressure of the voiding contractions and mean intermicturition interval in each group of rats ( n = 5). D Representative images of cystometrography results for each group of rats ( n = 5). E Masson staining results of bladder tissues in each group of rats. scale bar = 50 µm. F Immunofluorescence staining results for CD73 and VEGF in the bladder tissues of each group. Red indicates CD73, green indicates VEGF, and blue indicates DAPI. scale bar = 20 µm. G Quantitative analysis of smooth muscle content in the bladder (yellow square) of each group ( n = 3). H Quantitative analysis of immunofluorescence staining for CD73 and VEGF ( n = 3). Data presented as ±SD. * and # indicates P < 0.05, ** and ## indicates P < 0.01, and *** and ### indicates P < 0.001. Full-length blots/gels are presented in Supplementary Fig. .

Article Snippet: The samples were subjected to SDS-PAGE, and primary antibodies were used against CD73 (diluted 1:1000, Affinity, Ontario, Canada), VEGF (diluted 1:1000, proteintech, Chicago, IL, USA), SDF-1 (diluted 1:1000, proteintech, Chicago, IL, USA), NLRP3 (diluted 1:1000, Affinity, Ontario, Canada), p-AKT (diluted 1:1000, Affinity, Ontario, Canada), AKT (diluted 1:1000, Affinity, Ontario, Canada),caspase-1 (diluted 1:1000, Servicebio, Wuhan, Hubei, China), p-NFκB (diluted 1:1000, Cell Signaling Technology, Danvers, MA, USA), NF-κB (diluted 1:1000, Cell Signaling Technology, Danvers, MA, USA), and GAPDH (diluted 1:1000, Abcam, Cambridge, UK).

Techniques: Western Blot, Staining, Immunofluorescence

Journal: NPJ Regenerative Medicine

Article Title: CD73 overexpression in ADSCs accelerates bladder repair by regulating the NFκB/NLRP3/caspase-1 signaling axis in neurogenic bladder rats

doi: 10.1038/s41536-026-00454-1

Figure Lengend Snippet: A Immunofluorescence staining results of ADSCs in the bladder tissues of each group of rats. Red represents ADSCs, green represents βIII-tubulin, and blue represents DAPI. Scale bar = 20 µm. B Immunohistochemical results of CXCR4 in the bladder tissues of each group. The green arrows indicate regions of high CXCR4 expression. Scale bar = 50 µm. C Immunofluorescence staining results of CD73 and SDF-1 in the bladder tissues of each group. Red represents SDF-1, green represents CD73, and blue represents DAPI. Scale bar = 20 µm. D Quantitative results of ADSCs in the bladder tissues of each group ( n = 5). E Quantitative results of SDF-1 expression in the bladder tissues of each group ( n = 3). F In vitro, western blot results of SDF-1 in each group under conditioned medium. G Quantitative results of SDF-1 expression of each group ( n = 3). H Cell migration assay results of CD73⁺ ADSCs after SDF-1 gene knockdown. I Quantitative results of cell migration assays for ADSCs in each group ( n = 3). Data presented as ±SD. * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001. Full-length blots/gels are presented in Supplementary Fig. . Full-section IHC are presented in Supplementary Fig. .

Article Snippet: The samples were subjected to SDS-PAGE, and primary antibodies were used against CD73 (diluted 1:1000, Affinity, Ontario, Canada), VEGF (diluted 1:1000, proteintech, Chicago, IL, USA), SDF-1 (diluted 1:1000, proteintech, Chicago, IL, USA), NLRP3 (diluted 1:1000, Affinity, Ontario, Canada), p-AKT (diluted 1:1000, Affinity, Ontario, Canada), AKT (diluted 1:1000, Affinity, Ontario, Canada),caspase-1 (diluted 1:1000, Servicebio, Wuhan, Hubei, China), p-NFκB (diluted 1:1000, Cell Signaling Technology, Danvers, MA, USA), NF-κB (diluted 1:1000, Cell Signaling Technology, Danvers, MA, USA), and GAPDH (diluted 1:1000, Abcam, Cambridge, UK).

Techniques: Immunofluorescence, Staining, Immunohistochemical staining, Expressing, In Vitro, Western Blot, Cell Migration Assay, Knockdown, Migration

Journal: NPJ Regenerative Medicine

Article Title: CD73 overexpression in ADSCs accelerates bladder repair by regulating the NFκB/NLRP3/caspase-1 signaling axis in neurogenic bladder rats

doi: 10.1038/s41536-026-00454-1

Figure Lengend Snippet: CD73 activation upregulates VEGF expression, further stimulating the PI3K/AKT/mTOR pathway to enhance cell proliferation. Simultaneously, it inhibits NFκB phosphorylation, suppressing the NFκB/NLRP3/caspase-1 axis, thereby preventing apoptosis and reducing IL-1β and IL-6 levels. Moreover, activated CD73 increases SDF-1 expression, which interacts with its receptor CXCR4 to direct cell migration to damaged bladder tissue.

Article Snippet: The samples were subjected to SDS-PAGE, and primary antibodies were used against CD73 (diluted 1:1000, Affinity, Ontario, Canada), VEGF (diluted 1:1000, proteintech, Chicago, IL, USA), SDF-1 (diluted 1:1000, proteintech, Chicago, IL, USA), NLRP3 (diluted 1:1000, Affinity, Ontario, Canada), p-AKT (diluted 1:1000, Affinity, Ontario, Canada), AKT (diluted 1:1000, Affinity, Ontario, Canada),caspase-1 (diluted 1:1000, Servicebio, Wuhan, Hubei, China), p-NFκB (diluted 1:1000, Cell Signaling Technology, Danvers, MA, USA), NF-κB (diluted 1:1000, Cell Signaling Technology, Danvers, MA, USA), and GAPDH (diluted 1:1000, Abcam, Cambridge, UK).

Techniques: Activation Assay, Expressing, Phospho-proteomics, Migration

Journal: Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics

Article Title: RNA Interference of Biot2 Induces G₁ Phase Arrest and Apoptosis in Mouse Colorectal Cancer Cell Line

doi: 10.3727/096504014x14146137738583

Figure Lengend Snippet: Figure 2. Biot2 genes downregulated arrest of CT26 cell cycle. The effect of Biot2-shRNA on inducing G1 cell cycle arrest in CT26 cells. After treatment with Biot2-shRNA for 48 h, (A) the cell cycle distributions were determined by flow cytometry. (B) The protein expressions of cycle regulatory proteins and cyclin-dependent kinase cyclin D1, CDK2 and (C) the expression of several CKIs, p16, p21, and p27, were measured by Western blotting.

Article Snippet: Rabbit anti-CDK2 (1:1,000), rabbit anti-p16 (1:1,000), rabbit anti-p27 (1:1,000), and mouse anti-PCNA (1:1,000) were purchased from BOSTER (China).

Techniques: shRNA, Flow Cytometry, Expressing, Western Blot

Journal: Saudi Journal of Biological Sciences

Article Title: Effect of stachydrine hydrochloride to the prostate hyperplasia model in mice

doi: 10.1016/j.sjbs.2018.12.012

Figure Lengend Snippet: Effect of ACP, non PACP, PACP levels of the prostate hyperplasiamice model mice serum. (A): Level of ACP (U/L); (B): Level of Non- PACP (U/L); (C): Level of PACP (U/L). In A, B, C, “a” represents a significant difference ( P < 0.01) between the different administration groups compared with MG, and “b” represents there were significant differences between the different drug groups compared with MG ( P < 0.05), n = 10 mices/group.

Article Snippet: Stachydrine hydrochloride, Sichuan Institute of natural active ingredients, the content is more than 90%, Batch number: 20091212; Finasteride Capsules, Jiangsu Yabang Johnson Pharmaceutical Co Ltd, batch number: 080714; formaldehyde, Zhengzhou painI chemical reagent factory, Batch number: 20090401; Sodium Chloride Injection, Zheng Zhouyong and pharmaceutical Limited by Share Ltd, Batch number:20091002; Benzylpenicillin sodium injection, North China Pharmaceutical Limited by Share Ltd, Batch number: Y0903319; dihydrotestosterone (DHT), RD, batch number:20091216; total acid phosphatase (ACP), nonspecific acid phosphatase (non PACP), prostatic acid phosphatase (PACP), Nanjing Jiancheng Biological Engineering Institute, Batch number:20091228; bFGF, TGF-, EGF, beta 1 I IGF- kit were purchased from Wuhan boster Biological Engineering Co Ltd.

Techniques:

Journal: Histochemistry and Cell Biology

Article Title: Origin of renal myofibroblasts in the model of unilateral ureter obstruction in the rat

doi: 10.1007/s00418-008-0433-8

Figure Lengend Snippet: Renal cortex in sham-operated ( a ) and in ureter-ligeted kidneys ( b , c ). 3 μm cryostat sections; immunofluorescence; ecto-5′nucleotidase ( 5′NT )— red channel , alpha smooth muscle actin ( αSMA )— green channel ); chromatin staining by DAPI— blue channel . In controls ( a ) 5′NT strongly labels cells in the interstitium and the brush border of proximal tubules, αSMA labels exclusively smooth muscle cells in arterial vessels ( a ). In the interstitium of ureter-ligated kidneys ( b , c ) 5′NT staining decreases, whereas that of αSMA appears and becomes increasingly prominent. d – f interstitial fibroblast in ureter-ligated kidney after 2 days. 5′NT is distributed over the plasma membrane and cytoplasm in a granular manner, αSMA is apparent along the plasma membrane and in the cellular processes. Bar a – c ~100 μm, d – f ~10 μm

Article Snippet: Ecto-5′nucleotidase (5′NT) , Mouse , BD Biosciences, Franklin Lakes, NJ, USA.

Techniques: Immunofluorescence, Staining

Journal: Histochemistry and Cell Biology

Article Title: Origin of renal myofibroblasts in the model of unilateral ureter obstruction in the rat

doi: 10.1007/s00418-008-0433-8

Figure Lengend Snippet: Renal cortex after 4 days of ureter ligation; 3 μm cryostat sections; immunofluorescence; ecto-5′ nucleotidase ( 5′NT )— red channel , alpha smooth muscle actin ( αSMA )— green channel ); chromatin staining by DAPI— blue channel . a Merge of channels, b 5′NT, c αSMA; the framed area 1 comprises interstitium with high 5′NT and faint αSMA staining, adjacent to rather intact tubules, the framed area 2 comprises interstitium with strong αSMA and faint 5′NT staining, adjacent to collapsed tubules. Bar a–c ~100 μm; 1, 2: 10 μm

Article Snippet: Ecto-5′nucleotidase (5′NT) , Mouse , BD Biosciences, Franklin Lakes, NJ, USA.

Techniques: Ligation, Immunofluorescence, Staining

Journal: Histochemistry and Cell Biology

Article Title: Origin of renal myofibroblasts in the model of unilateral ureter obstruction in the rat

doi: 10.1007/s00418-008-0433-8

Figure Lengend Snippet: Mitotic cells in the peritubular interstitium in ureter-ligated kidneys. 3 μm cryostat sections, immonofluorescence; αSMA— green channel ; 5′NT, or phopho-S6 kinase (p-S6 K)— red channel , chromatin staining by DAPI— blue channel . The mitotic cell in a heavily expresses 5′NT and faintly αSMA, the adjacent cell shows strong αSMA and faint 5′NT staining. The two mitotic cells in B display strong staining for αSMA along the plasma membrane and in the cell processes, 5′NT staining appears granular and is seen over the cytoplasm. The mitotic cell in C shows strong αSMA along the plasma membrane, in the cell processes and in a reticulated manner also in the cytoplasm; p-S6 K is up regulated in the cytoplasm of mitotic cells, whereas in quiescent cells the nucleus shows weak staining. Bar ~10 μm

Article Snippet: Ecto-5′nucleotidase (5′NT) , Mouse , BD Biosciences, Franklin Lakes, NJ, USA.

Techniques: Staining

Journal: Histochemistry and Cell Biology

Article Title: Origin of renal myofibroblasts in the model of unilateral ureter obstruction in the rat

doi: 10.1007/s00418-008-0433-8

Figure Lengend Snippet: FSP1/S100A4-positive cells in the renal cortex. 3 μm cryostat sections, immonofluorescence; 5′NT— red channel ; FSP 1/S100A4— green channel . In controls a very few FSP1/S100A4-positive cells are seen in the peritubular interstitium among the 5’NT-positive interstitial cells; the abundance of FSP1/S100A4 positive cells in the interstitial spaces increases at day 2 and 3 after ureteral obstruction; a few are found in glomeruli ( G ); arterioles ( arrow ) are weakly labeled by the antibody to FSP1/S100A4, tubular epithelia are negative for FSP1/S100A4. Bar ~100 μm

Article Snippet: Ecto-5′nucleotidase (5′NT) , Mouse , BD Biosciences, Franklin Lakes, NJ, USA.

Techniques: Labeling

Journal: PLoS ONE

Article Title: Influence of Murine Mesenchymal Stem Cells on Proliferation, Phenotype, Vitality, and Cytotoxicity of Murine Cytokine-Induced Killer Cells in Coculture

doi: 10.1371/journal.pone.0088115

Figure Lengend Snippet: MSCs were harvested and analysed with flow cytometry for a panel of MSC-characterizing markers on day 7. Thus, the presented phenotype resembles that of MSCs used in the cocultures. MSCs were negative for CD3, CD31, CD34, and CD90. Positive signals were measured for CD44, CD73, and CD166. Positive as well as negative signals were detected for CD45.

Article Snippet: In addition, CD34-PE (Caltaq, 1∶100), CD44-AF488 (Biozol, 1∶250), CD73-PE (Biozol, 1∶100), CD90 (Thy1.2)-FITC (Miltenyi, 1∶100), and CD166 (Antikörper-online, 1∶100).

Techniques: Flow Cytometry