Journal: Molecular Imaging

Article Title: PET Study of Sphingosine-1-phosphate Receptor 1 Expression in Response to S. aureus Infection

doi: 10.1155/2021/9982020

Figure Lengend Snippet: MicroPET imaging of S1PR1 activity in S aureus -infected mice. (a) Radiosynthesis of S1PR1-specific radiotracer, [ 18 F]TZ4877; (b) representative sagittal microPET images of [ 18 F]TZ4877 in mice. Comparing with sham mice, the tracer uptake was significantly higher in the infected mice, and the increased uptake of the tracer showed S aureus dose dependent; (c) the tracer uptake in the brain was quantified; time-activity curves showed that the tracer uptake in infected mice was significantly higher than mice without infections; (d) the average tracer uptake in the brain from 30 to 50 min of the PET scan showed a dose-dependent manner. Data represent the mean ± SEM, n = 3 for each group.

Article Snippet: After washing in PBS, all sections were then incubated with rabbit anti-S1PR1 (Alomone, Israel) antibody overnight at 4°C and then washed and incubated with ImmPRESS HRP Horse anti-rabbit polymer for 1 hour at room temperature and developed with ImmPACT DAB (Vector Laboratories, Burlingame, CA).

Techniques: Imaging, Activity Assay, Infection

Journal: Molecular Imaging

Article Title: PET Study of Sphingosine-1-phosphate Receptor 1 Expression in Response to S. aureus Infection

doi: 10.1155/2021/9982020

Figure Lengend Snippet: Biodistribution (%ID/g, mean ± SEM) of S1PR1-specific [ 18 F]TZ4877 in Balb/c mice ( n = 4).

Article Snippet: After washing in PBS, all sections were then incubated with rabbit anti-S1PR1 (Alomone, Israel) antibody overnight at 4°C and then washed and incubated with ImmPRESS HRP Horse anti-rabbit polymer for 1 hour at room temperature and developed with ImmPACT DAB (Vector Laboratories, Burlingame, CA).

Techniques: Mouse Assay

Journal: Molecular Imaging

Article Title: PET Study of Sphingosine-1-phosphate Receptor 1 Expression in Response to S. aureus Infection

doi: 10.1155/2021/9982020

Figure Lengend Snippet: Biodistribution of S1PR1-specific [ 18 F]TZ4877 in sham, infected, and infected with treatments mice ( n = 4).

Article Snippet: After washing in PBS, all sections were then incubated with rabbit anti-S1PR1 (Alomone, Israel) antibody overnight at 4°C and then washed and incubated with ImmPRESS HRP Horse anti-rabbit polymer for 1 hour at room temperature and developed with ImmPACT DAB (Vector Laboratories, Burlingame, CA).

Techniques: Infection, Mouse Assay

Journal: Molecular Imaging

Article Title: PET Study of Sphingosine-1-phosphate Receptor 1 Expression in Response to S. aureus Infection

doi: 10.1155/2021/9982020

Figure Lengend Snippet: MicroPET imaging of S1PR1 activity in S aureus -infected mice. (a) Representative sagittal microPET images of [ 18 F]TZ4877 in the hind limb of mice. The tracer uptake was relatively low in the hind limb muscle with a SUV of ~1.5 in sham mice. Comparing with sham mice, the tracer uptake was significantly higher in the hind limb of infected mice; (b) time-activity curves showed that the tracer uptake in infected mice was significantly higher than sham mice; (c) the average tracer uptake in the hind limb muscle from 30 to 50 min of the PET scan showed a ~39% increase of SUV in infected mice with a P value of 0.0082. Data represent the mean ± SEM, n = 3 for each group.

Article Snippet: After washing in PBS, all sections were then incubated with rabbit anti-S1PR1 (Alomone, Israel) antibody overnight at 4°C and then washed and incubated with ImmPRESS HRP Horse anti-rabbit polymer for 1 hour at room temperature and developed with ImmPACT DAB (Vector Laboratories, Burlingame, CA).

Techniques: Imaging, Activity Assay, Infection

Journal: Molecular Imaging

Article Title: PET Study of Sphingosine-1-phosphate Receptor 1 Expression in Response to S. aureus Infection

doi: 10.1155/2021/9982020

Figure Lengend Snippet: PET measurements of S1PR1-specific [ 18 F]TZ4877 in S aureus -infected and sham mice.

Article Snippet: After washing in PBS, all sections were then incubated with rabbit anti-S1PR1 (Alomone, Israel) antibody overnight at 4°C and then washed and incubated with ImmPRESS HRP Horse anti-rabbit polymer for 1 hour at room temperature and developed with ImmPACT DAB (Vector Laboratories, Burlingame, CA).

Techniques: Infection

Journal: Molecular Imaging

Article Title: PET Study of Sphingosine-1-phosphate Receptor 1 Expression in Response to S. aureus Infection

doi: 10.1155/2021/9982020

Figure Lengend Snippet: Immunohistochemistry analysis of S1PR1 in hind limb muscle of sham and S aureus -infected mice. S1PR1 was significantly upregulated in the muscle of infected mice (red arrow) comparing with sham mice (green arrow), scale bar = 100 μ m.

Article Snippet: After washing in PBS, all sections were then incubated with rabbit anti-S1PR1 (Alomone, Israel) antibody overnight at 4°C and then washed and incubated with ImmPRESS HRP Horse anti-rabbit polymer for 1 hour at room temperature and developed with ImmPACT DAB (Vector Laboratories, Burlingame, CA).

Techniques: Immunohistochemistry, Infection

Journal: Acta pharmaceutica Sinica. B

Article Title: S1PR1 serves as a viable drug target against pulmonary fibrosis by increasing the integrity of the endothelial barrier of the lung.

doi: 10.1016/j.apsb.2022.10.006

Figure Lengend Snippet: Figure 1 S1pr1 expressed in ECs of lung tissues and downregulated in IPF patients. The distribution of S1pr1 in human lung tissues (A, B). (A) Clusters of human lung tissues were identified as four cell types, including endothelial, epithelial, immune, and mesenchymal cells. (B) S1pr1 was highly expressed in endothelial (yellow) and immune cells (light green). The expression of S1pr1 in human lung tissues (C, D). (C) The RNA expression of S1pr1 in lung tissues between nonfibrotic individuals (n Z 10, Tobacco Users, 8 of 10) and IPF patients (n Z 12) were analyzed. (D) The RNA expression of S1pr1 in ECs between healthy individuals (n Z 10) and IPF patients (n Z 12) were measured. ****P < 0.0001 vs. control. (E) S1pr1 was highly expressed in lung ECs and immune cells (n Z 7). (F) Box and whisker plot showed the expression of S1pr1 in lung. (G) Real-time qPCR analysis of S1pr1 expression in primary fibroblasts, endothelial and epithelial cells of mouse lung tissues (n Z 3). Bars represent mean SEM, ****P < 0.0001 vs. fibroblasts.

Article Snippet: S1pr1 conditional genetically targeted (S1pr1þ/ ) mice and TekCreERT2 mice were generated by Cyagen Biosciences (Suzhou, China).

Techniques: Expressing, RNA Expression, Control, Whisker Assay

Journal: Acta pharmaceutica Sinica. B

Article Title: S1PR1 serves as a viable drug target against pulmonary fibrosis by increasing the integrity of the endothelial barrier of the lung.

doi: 10.1016/j.apsb.2022.10.006

Figure Lengend Snippet: Figure 2 S1pr1 deficiency in ECs worsened bleomycin-induced injury and fibrosis in mouse lungs. (A) S1pr1f/f mice were generated by gene targeting and they were crossed with the Tek-CreERT2 mice to obtain endothelial-specific S1pr1 knockout mice, which were named as S1pr1þ/

Article Snippet: S1pr1 conditional genetically targeted (S1pr1þ/ ) mice and TekCreERT2 mice were generated by Cyagen Biosciences (Suzhou, China).

Techniques: Generated, Knock-Out

Journal: Acta pharmaceutica Sinica. B

Article Title: S1PR1 serves as a viable drug target against pulmonary fibrosis by increasing the integrity of the endothelial barrier of the lung.

doi: 10.1016/j.apsb.2022.10.006

Figure Lengend Snippet: Figure 3 S1pr1 deficiency of ECs destroyed the integrity of vascular barrier and increased inflammation and fibrosis. (A) Heatmap of mRNA expression of CD31þ ECs of the lung (left) and volcano map of DEGs (middle). GO enrichment analysis (right) (n Z 3). (B) GSEA and heatmap of tight junction-related genes and heatmap of tight junction genes. (C) Western blot analysis of ZO-1 in the lung section (top). Statistical analysis of the expression of ZO-1 (left bottom). ZO-1 mRNA expression from transcriptome analysis (right bottom) (n Z 3). (D) Statistical analysis of CD45þ cell percentage of total cells (left) and MFI of a-SMA near CD31þ cells (right) (n Z 3). (E) Representative results for mIHC staining of CD45 in the lung sections from S1pr1þ/ con and Cre con mice. Nuclei were stained blue by DAPI, and the images were obtained at an original magnification of 20. (F) Representative results for mIHC staining of CD31 and a-SMA in the lung sections from S1pr1þ/ con and Cre con mice. Nuclei were stained blue by DAPI, and the images were obtained at an original magnification of 20. Bars represent mean SEM, *P < 0.05 and **P < 0.01 vs. Cre con group.

Article Snippet: S1pr1 conditional genetically targeted (S1pr1þ/ ) mice and TekCreERT2 mice were generated by Cyagen Biosciences (Suzhou, China).

Techniques: Expressing, Western Blot, Staining

Journal: Acta pharmaceutica Sinica. B

Article Title: S1PR1 serves as a viable drug target against pulmonary fibrosis by increasing the integrity of the endothelial barrier of the lung.

doi: 10.1016/j.apsb.2022.10.006

Figure Lengend Snippet: Figure 4 S1pr1 deficiency of ECs accelerated BLM-induced fibrosis in the lung. (A) Heatmap of mRNA expression of CD31þ ECs of the lung (left).VolcanomapofDEGs(middle).GOenrichmentanalysis(right)(B)GSEAwasperformed(nZ4).(C)Heatmapanalysisofcollagen(left),ECM (middle), and tight junction (right) related genes (n Z 4). (D) Representative results for mIHC staining of CD45þ in the lung sections from S1pr1þ/

Article Snippet: S1pr1 conditional genetically targeted (S1pr1þ/ ) mice and TekCreERT2 mice were generated by Cyagen Biosciences (Suzhou, China).

Techniques: Expressing, Staining

Journal: Acta pharmaceutica Sinica. B

Article Title: S1PR1 serves as a viable drug target against pulmonary fibrosis by increasing the integrity of the endothelial barrier of the lung.

doi: 10.1016/j.apsb.2022.10.006

Figure Lengend Snippet: Figure 5 Activation of S1PR1 by selective S1PR1 agonist, IMMH002, ameliorated mouse PF induced by BLM. (A) Survival curve of S1PR1 agonists on BLM-induced mouse fibrosis model. (B) Calculated lung index of animal model. Lung index Z weight of lung (g)/body weight (g) 1000 (n Z 7e10). (C) Lung function detected by pulmonary function test, including IC, Crs, Ras, Ers, Cst, G, H, and Est (n Z 3e5). (D) Histological analysis of the severity of lung fibrosis after BLM induction. Representative images for HE (top), Masson staining (bottom). Images were obtained at 200 magnification. (E) Statistical analysis of pathology score (left) and fibrosis score (right) (n Z 7e10). (F) Western blot analysis of ZO-1, E-cadherin, Snail, fibronectin, and b-actin (n Z 3). (G) Statistical analysis of Western blot. Bars represent mean SEM, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 vs. Model, #P < 0.05, ##P < 0.01 and ####P < 0.0001 vs. Sham.

Article Snippet: S1pr1 conditional genetically targeted (S1pr1þ/ ) mice and TekCreERT2 mice were generated by Cyagen Biosciences (Suzhou, China).

Techniques: Activation Assay, Animal Model, Staining, Western Blot

Journal: Acta pharmaceutica Sinica. B

Article Title: S1PR1 serves as a viable drug target against pulmonary fibrosis by increasing the integrity of the endothelial barrier of the lung.

doi: 10.1016/j.apsb.2022.10.006

Figure Lengend Snippet: Figure 6 The selective S1PR1 agonist, IMMH002, increase endothelial barrier. (A) Immunofluorescence staining of ZO-1 in HUVECs. Nuclei were stained blue by DAPI, and the images were obtained at an original magnification of 40 . (B) Statistical analysis of the MFI of ZO-1. (C) Relative EC permeability Z Fluorescence value of the lower chamber/Fluorescence value of the upper chamber and standardized by control groups. (D) TEER measurements were performed by the Millicell voltammeter. TEER (U$cm2) Z Resistance (U) 0.333 (R Z 3.25 mm, S Z 0.333 cm2) and standardized by control groups. Bars represent mean SEM, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 vs. PBS with thrombin; ###P < 0.001 and ####P < 0.0001 vs. Control. (E) KM mice were administrated with IMMH002 and DXM for 7 continuous days. Thereafter, mice were injected with 1% acetic acid to form a peritoneal leakage model. Peritoneal lavage fluid absorbance was measured at 590 nm (n Z 7e9). Bars represent mean SEM, *P < 0.05 vs. Control. (F) Visualization of endothelial cytoskeletal rearrangement. Nuclei were stained blue by DAPI, and the images were obtained at an original magnification of 600 . (G) Immunofluorescence staining of CD31 and ZO-1 in mouse lung sections. Nuclei were stained blue by DAPI, and the images were obtained at an original magnification of 600 .

Article Snippet: S1pr1 conditional genetically targeted (S1pr1þ/ ) mice and TekCreERT2 mice were generated by Cyagen Biosciences (Suzhou, China).

Techniques: Staining, Permeability, Fluorescence, Control, Injection

Journal: Acta pharmaceutica Sinica. B

Article Title: S1PR1 serves as a viable drug target against pulmonary fibrosis by increasing the integrity of the endothelial barrier of the lung.

doi: 10.1016/j.apsb.2022.10.006

Figure Lengend Snippet: Figure 7 An image illustrating how endothelial barrier was disrupted by S1pr1 deficiency. S1PR1 agonist, IMMH002, could ameliorate PF by enhancing endothelial tight junctions.

Article Snippet: S1pr1 conditional genetically targeted (S1pr1þ/ ) mice and TekCreERT2 mice were generated by Cyagen Biosciences (Suzhou, China).

Techniques:

Journal: Journal of Neuroinflammation

Article Title: Cry 2 deficiency leads to cognitive impairment through the microbiota-gut-brain axis mediated S1P/NLRP3/IL-1β pathway in mice

doi: 10.1186/s12974-026-03706-5

Figure Lengend Snippet: Gut microbiota induces sphingolipid pathway disturbance in the peripheral and brain. ( A ) Representative metabolites classified and counted based on their chemical taxonomy in shScr and shCry2 mice. The size of the pie chart corresponded to the relative abundance levels of metabolites, (n = 10 mice/group)., (B) Volcano plot of differentially expressed metabolites from shScr versus shCry2 mice. Blue, red, and grey represented downregulated, upregulated, and no significantly expressed metabolites, respectively, (n = 10 mice/group)., ( C ) Unsupervised hierarchical clustering of differentially expressed metabolites from shScr and shCry2 mice, (n = 10 mice/group)., ( D ) Pathway enrichment analysis of differentially expressed metabolites between shScr and shCry2 mice, (n = 10 mice/group)., ( E ) Representative Western blot and quantification of Sphk1 and S1PR1 in the hippocampus of shScr and shCry2 mice., ( F ) LC/MS analysis of Cer(18:1/18:0) and Cer(18:2/18:0) in shScr and shCry2 mice., ( G ) Linear regression analyses between sphingosine and Akkermansia, (left plot) and Ruminococcaceae, (right plot) expression levels from data integrated from 16 S rRNA sequencing and metabolomic sequencing. All data were expressed as mean ± SEM. Data were considered significant if * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Means were compared using the Student’s t-test in panel, ( E , F ). shScr: shScramble

Article Snippet: Then the membranes were incubated overnight at 4 °C with the primary antibodies [Tau5 (1:1000, ab80579, Abcam), Tau1 (1:1000, MAB3420, MilliporeSigma), pS396-tau (1:5000, ab109390, Abcam), pT231-tau (1:5000, ab151559, Abcam), pT181-tau (1:5000, ab254409, Abcam), pT217-tau (1:2000, 44–744, Invitrogen), Occludin (1:1000, ab216327, Abcam), ZO-1 (1:1000, ab307799, Abcam), Sphk1 (1:1000, 10670-1-AP, Proteintech), CRY2 (1:1000, 13997-1-AP, Proteintech), S1PR1 (1:1000, 55133-1-AP, Proteintech), β-actin (1:5000, 66009-1-Ig, Proteintech), NLRP3 (1:1000, ab263899, Abcam), IL-1β (1:1000, ab234437, Abcam)] diluted in 1×TBST and washed 10 min three times.

Techniques: Western Blot, Liquid Chromatography with Mass Spectroscopy, Expressing, Sequencing, Metabolomic, IF-P

Journal: Journal of Neuroinflammation

Article Title: Cry 2 deficiency leads to cognitive impairment through the microbiota-gut-brain axis mediated S1P/NLRP3/IL-1β pathway in mice

doi: 10.1186/s12974-026-03706-5

Figure Lengend Snippet: FTY720 ameliorated the cognitive decline, impairment of BBB and tau pathology. ( A ) An overview of the experimental design. After receiving stereotaxic brain injections, three-month-old mice were housed for six weeks, followed by intraperitoneal injections of saline or FTY720 for two weeks. Behavioral and molecular biology tests were then conducted., ( B - E ) Escape latency of the shScr-saline, shScr-FTY720, shCry2-saline, and shCry2-FTY720 mice for 5 consecutive days in the MWM test, ( B ). Percentage of time spent in the target quadrant in the spatial probe trail of the MWM test, ( C ). Number of platform crossings in the spatial probe trail of the MWM test, ( D ). Representative heatmaps of the swimming path in the spatial probe trail of day 6 in the MWM test, ( E ), (n = 11–12 mice/group)., ( F - H ) Representative Western blot and quantification of Occludin, ZO-1, ( F ), Sphk1, S1PR1, ( G ), Tau1, pS396-tau, Tau5, and pT231-tau, ( H ) in the hippocampus of shScr-saline, shScr-FTY720, shCry2-saline, and shCry2-FTY720 mice., ( I , J ) Representative immunofluorescence staining and quantification of pS396-tau, (red) in the hippocampus of shScr-saline, shScr-FTY720, shCry2-saline, and shCry2-FTY720 mice. Magnification × 10. Scale bar = 100 μm. All data were expressed as mean ± SEM. Data were considered significant if *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Means were compared using a repeated measures 2-way ANOVA test with Bonferroni post hoc comparison in panel, ( B ) and one-way ANOVA, ( C - D , F - I ). FI: Fluorescence intensity. shScr: shScramble

Article Snippet: Then the membranes were incubated overnight at 4 °C with the primary antibodies [Tau5 (1:1000, ab80579, Abcam), Tau1 (1:1000, MAB3420, MilliporeSigma), pS396-tau (1:5000, ab109390, Abcam), pT231-tau (1:5000, ab151559, Abcam), pT181-tau (1:5000, ab254409, Abcam), pT217-tau (1:2000, 44–744, Invitrogen), Occludin (1:1000, ab216327, Abcam), ZO-1 (1:1000, ab307799, Abcam), Sphk1 (1:1000, 10670-1-AP, Proteintech), CRY2 (1:1000, 13997-1-AP, Proteintech), S1PR1 (1:1000, 55133-1-AP, Proteintech), β-actin (1:5000, 66009-1-Ig, Proteintech), NLRP3 (1:1000, ab263899, Abcam), IL-1β (1:1000, ab234437, Abcam)] diluted in 1×TBST and washed 10 min three times.

Techniques: Saline, Western Blot, Immunofluorescence, Staining, IF-P, Comparison, Fluorescence

Journal: eLife

Article Title: A novel role for lipoxin A 4 in driving a lymph node–eye axis that controls autoimmunity to the neuroretina

doi: 10.7554/eLife.51102

Figure Lengend Snippet: All tissues were collected from mice immunized with 150 μg of IRBP 651-670 . ( A–B ) Heat maps showing gene expression of inflammation and cell trafficking markers in the eyes and inguinal lymph nodes harvested from EAU-challenged mice treated with vehicle or LXA 4 . ( A ) Inflammation markers and ( B ) migration markers analyzed by nanoString. Each sample was pooled from 3 to 4 mice on day 16 post-immunization. ( C ) S1pr1 expression relative to β-actin from eyes and various lymph nodes of mice treated with vehicle or LXA 4 harvested on day 16 post-immunization, same samples as panels A and B. ( D ) Fold change in S1pr1 expression of CD4 + T cells isolated from WT and Alox5 -/- on day 14 post-immunization, n = 11 from three experiments combined. ( E ) Transwell migration assay of CD4 + T cells isolated from immunized WT and Alox5 -/- on day 13 post-immunization. CD4 + T cells were pre-stimulated with anti-CD3 and anti-CD28 antibodies for 18 hr and transferred to transwell culture plates and incubated for 4 hr in the absence or presence of CCL19 and CCL21, n = 12 per group. One representative experiment of 4 showing the same trend. **p<0.01, unpaired Welch’s t test. ( F–G ) Representative flow plots of CCR7 and S1PR1 expression on in vitro anti-CD3 and anti-CD28 stimulated CD4 + T cells treated with vehicle control or 10 nM, 100 nM, 1 μM, and 2 μM of LXA 4 . Cells were gated on live single cells then CD4 + cells. Gray overlays indicate fluorescence minus one controls. Showing one representative experiment out of three.

Article Snippet: Alox5 : Mm01182747_m1, Alox15 : Mm00507789_m1, Fpr2 : Mm00484464_s1, S1pr1 : Mm00514644_m1.

Techniques: Gene Expression, Migration, Expressing, Isolation, Transwell Migration Assay, Incubation, In Vitro, Control, Fluorescence

Journal: Fluids and Barriers of the CNS

Article Title: APPswe mutation causes functional deficits in endothelial cells generated by transient ETV2 overexpression in human iPSCs

doi: 10.1186/s12987-025-00728-8

Figure Lengend Snippet: ETV2-ECs Respond to angiogenic stimuli by forming vessel-like structures in both 2D and 3D models. ( A ) Relative mRNA levels of KDR and S1PR1 in hiPSC-pericytes, S-ECs, ETV2-ECs, and hiPSCs., shown as fold change relative to GAPDH. ( B ) Representative 2D tube formation images of S and ETV2-ECs with and without sprouting mix, 6 h post-exposure. Scale bars: 800 μm. ( C ) Quantification of tube formation metrics: number of master segments, master segment lengths, mesh counts, and mesh areas. ( D ) Representative 3D vessel formation images showing sprouting ends formed by S and ETV2-ECs with and without sprouting mix, 24 h post-exposure. Scale bars: 100 μm. ( E ) Quantification of sprouting events, expressed as the number of sprouts per 100 μm of spheroid perimeter in S and ETV2-EC spheroids. Dot plots represent the average of technical replicates for each differentiation batch, color-coded by hiPSC line. Data are presented as mean ± SD. Statistical analysis was performed using two-way ANOVA with Bonferroni’s multiple comparison test. Significance levels are denoted as * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: S1PR1 , Hs05021992_s1 , TaqMan, Thermo Fisher Scientific.

Techniques: Comparison

Journal: Fluids and Barriers of the CNS

Article Title: APPswe mutation causes functional deficits in endothelial cells generated by transient ETV2 overexpression in human iPSCs

doi: 10.1186/s12987-025-00728-8

Figure Lengend Snippet: Reduced vessel-like structure formation in APPswe ECs following sprouting mix stimulation. (A) Representative 2D tube formation images of control and APPswe ECs with and without sprouting mix, 6 h post-exposure. Scale bars: 800 μm. (B) Quantification of tube formation metrics: number of master segments, master segment lengths, mesh counts, and mesh areas. (C) Representative 3D vessel formation images showing sprouting ends formed by control and APPswe ECs with and without sprouting mix, 24 h post-exposure. Scale bars: 100 μm. (D) Quantification of sprouting events, expressed as the number of sprouts per 100 μm of spheroid perimeter in control and APPswe EC spheroids. (E) Relative mRNA levels of KDR and S1PR1 in control and APPswe ECs, normalized to GAPDH and expressed as fold change. Dot plots represent the average of technical replicates for each differentiation batch, color-coded by hiPSC line. Data are presented as mean ± SD. Statistical analysis was performed using two-way ANOVA with Bonferroni’s multiple comparison test. Significance levels are denoted as *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001

Article Snippet: S1PR1 , Hs05021992_s1 , TaqMan, Thermo Fisher Scientific.

Techniques: Control, Comparison

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: miRNA-503 inhibition exerts anticancer effects and reduces tumor growth in mesothelioma

doi: 10.1186/s13046-025-03283-0

Figure Lengend Snippet: Histological analysis of miR-503 target genes and transcriptomic analysis of miR-503 inhibition. A Evaluation of the expression of BTG1, CCNG1, EDG1 and TIMP2, in normal pleural (PL) and pleural mesothelioma tissues (EPM epithelial istotype; SPM, sacomatoid istotype). Yellow Scale bar = 100 μm. Data shown are mean ± S.D. of five independent experiments. B IHC analysis of the putative miR-503 target genes on mice tumors treated with LNPs-anti-miR-503 or empty LNPs (original magnification 20X). A and B confirm the in vitro results. C Transcriptome analysis in MSTO cells after miR-503 inhibition detected about 500 deregulated genes (red upregulated, green downregulated). D Expression analysis of MM biomarkers deregulated by inhibition of miR-503 in MM cells. E q-PCR analysis in human MM samples of the miR-503 MM biomarkers confirms the inverse expression only for CXCL8, SERPINE1 and SPP. F IHC analysis of the miR-503 deregulated MM biomarkers on mice tumors treated with LNPs-anti-miR-503 or empty LNPs (original magnification 20X). For each IHC panel bar plots report the score for the samples analysed. Data presented as the mean ± SD of at least three independent experiments ( n = 3) for the in vitro experiments while for the in vivo analysis numbers of samples are reported in Methods. * p < 0.05, ** p < 0.01 *** p < 0.001 **** p < 0.0001 vs. control

Article Snippet: The primary antibodies used were: Rabbit anti-human Ki67 (DAKO Agilent, Santa Clara, CA USA), Rabbit anti-human BTG1 (orb35408 Biorbyt, Durham, NC, USA), Rabbit anti-human CCNG1 (orb167206 Biorbyt), Rabbit anti-human S1PR1 (EDG1) (orb350684 Biorbyt), Rabbit anti-human TIMP2 (orb543218 Biorbyt), Rabbit anti-human CXCL8 (17038-1-AP Proteintech Rosemont, IL, USA), Rabbit anti-human Osteopontin, (SPP 20416-1-AP Proteintech), Rabbit anti-human SERPINE1 (PAI-1 #49536 Cell Signaling, Danvers, MA, USA).

Techniques: Inhibition, Expressing, In Vitro, In Vivo, Control