Journal: Discover. Oncology

Article Title: SAA1 regulated by S1P/S1PR1 promotes the progression of ESCC via β-catenin activation

doi: 10.1007/s12672-024-00923-3

Figure Lengend Snippet: siRNA sequences

Article Snippet: After 12 h of starvation, the cells were treated with VPC23019 (S1PR1/S1PR3 antagonist, Sigma, USA) for 30 min, followed by stimulation with S1P.

Techniques:

Journal: Discover. Oncology

Article Title: SAA1 regulated by S1P/S1PR1 promotes the progression of ESCC via β-catenin activation

doi: 10.1007/s12672-024-00923-3

Figure Lengend Snippet: RT qPCR primer sequences

Article Snippet: After 12 h of starvation, the cells were treated with VPC23019 (S1PR1/S1PR3 antagonist, Sigma, USA) for 30 min, followed by stimulation with S1P.

Techniques: Quantitative RT-PCR

Journal: Discover. Oncology

Article Title: SAA1 regulated by S1P/S1PR1 promotes the progression of ESCC via β-catenin activation

doi: 10.1007/s12672-024-00923-3

Figure Lengend Snippet: S1P/S1PR1 increases SAA1 expression levels and β-catenin phosphorylation at Ser675 in ESCC cells. A ESCC cells were stimulated with different concentrations of S1P (0, 10, 100, and 1000 nmol/L) for 24 h. SAA1, total β-catenin and pSer675-β-catenin protein levels were detected by western blotting. B ESCC cells were treated with VPC23019 (1000 nmol/L) and then stimulated with S1P (1000 nmol/L) for 24 h. The levels of SAA1, total β-catenin and pSer675-β-catenin were detected by western blotting. C The expression of S1PR1 and S1PR3 proteins in HEEC cells and two ESCC cell lines was detected by western blotting. D The efficiency of S1PR1 overexpression was determined by RT‒qPCR and western blotting. E The efficiency of S1PR1 knockdown was determined by RT‒qPCR and western blot. F The protein levels of SAA1, total β-catenin and pSer675-β-catenin were detected by western blot after S1PR1 overexpression and knockdown in ESCC cells. **p < 0.01, ****p < 0.0001

Article Snippet: After 12 h of starvation, the cells were treated with VPC23019 (S1PR1/S1PR3 antagonist, Sigma, USA) for 30 min, followed by stimulation with S1P.

Techniques: Expressing, Western Blot, Over Expression

Journal: Heliyon

Article Title: Differences in collateral vessel formation after experimental retinal vein occlusion in spontaneously hypertensive rats and wild-type rats

doi: 10.1016/j.heliyon.2024.e27160

Figure Lengend Snippet: a): Immunofluorescence imaging of retinal flat mounts in Wild-type Wister rats (WWRs): occluded area (a–f) and non-occluded area (g–l). a, g: fluorescein isothiocyanate - labeled dextran (FITC/dextran) perfused retinas at 6 h after photocoagulation (PC). The red X indicates the site of vein occlusion. b, h: Immunohistochemical staining of sphingosine 1-phosphate receptor 1 (S1PR1) at 6 h after PC. c, I: Merged image from a, g and b, h. d, j: Enlarged view inside the yellow rectangle seen in a, g. e, k: Enlarged image from b, h showing enhanced S1PR1 expression along the blood vessels. f, l: Merged image of those from d, j and e, k. In b, h, enhanced S1PR1 expression was detected along the retinal vein occlusion. In e, k, enhanced S1PR1 expression was detected along the blood vessels. b): Immunofluorescence imaging of retinal flat mounts in spontaneously hypertensive rats (SHRs): occluded area (m–r) and non-occluded area (s–x): m, s: FITC/dextran-perfused retinas at 6 h after PC. The red X indicates the site of vein occlusion. n, t: Immunohistochemical staining of S1PR1 at 6 h after PC. O, u: Merged image of those from m, s and n, t. p, v: Enlarged view of the yellow rectangle seen in m, s. q, w: Enlarged image of that from n, t. r, x: Merged image of those from p, v and q, w. In n, t, enhanced S1PR1 expression was detected along the retinal vein occlusion. In q, w, enhanced S1PR1 expression was detected along the blood vessels. c): Immunofluorescence imaging of retinal flat mounts in WWRs as a negative control (NC):in the occluded (NC1–NC6) and non-occluded (NC7–NC12) areas: NC1, NC7: FITC/dextran-perfused retinas at 6 h after PC. The red X indicates the site of vein occlusion. NC2, NC8: Immunohistochemical staining of S1PR1 at 6 h after PC. NC3, NC9: Merged image of the images from NC1, NC7 and NC2, NC8. NC4, NC10: Enlarged view of the yellow rectangle in NC1, NC7.NC5, NC11: Enlarged image from NC2, NC8 showing enhanced S1PR1 expression along the blood vessels. NC6, NC12: Merged image of the image from NC4, NC10 and NC5, NC11.

Article Snippet: The WWRs and SHRs were each divided into four groups: control rats receiving no drug administration, rats treated with vehicle (dimethyl sulfoxide [DMSO]) only, rats treated with the S1PR1 agonist SEW2871 (SEW, Cayman Chemical, Ann Arbor, MI, USA), and rats treated with the S1PR1 antagonist VPC23019 (VPC, Cayman Chemical).

Techniques: Immunofluorescence, Imaging, Labeling, Immunohistochemical staining, Staining, Expressing, Negative Control

Journal: Heliyon

Article Title: Differences in collateral vessel formation after experimental retinal vein occlusion in spontaneously hypertensive rats and wild-type rats

doi: 10.1016/j.heliyon.2024.e27160

Figure Lengend Snippet: a): nitric oxide synthase 3 (NOS3) expression quantified by qRT-PCR in the whole retina of Wild-type Wister rats (WWRs) and spontaneously hypertensive rats (SHRs) before photocoagulation (PC) and 6 h after PC. The Y-axis indicate these are relative NOS3/gapdh to untreated control retina values. In both WWRs and SHRs, the expression of NOS3 was significantly higher at 6 h after PC than before PC (p < 0.001, Tukey–Kramer post-hoc test). Before the PC procedure, there was no significant difference in NOS3 expression between WWRs and SHRs. However, at 6 h after PC, NOS3 expression was predominantly higher in WWRs than SHRs (1.3 fold, p = 0.02, Tukey–Kramer post-hoc test). b): The number of PC shots required for vein occlusion before and after each drug administration. The WWRs and SHRs were each divided into four groups: control rats receiving no drug administration, rats treated with vehicle (dimethyl sulfoxide [DMSO]), rats treated with the S1PR1 agonist SEW2871, and rats treated with the S1PR1 antagonist VPC23019. The number of PC shots required to achieve retinal vein occlusion was higher in the SEW-treated group than control group in both WWRs and SHRs (p < 0.001, Tukey–Kramer post-hoc test). Specifically, in the SEW-treated WWRs, the three eyes that could be occluded required significantly more laser shots compared with any of the other groups (p < 0.001, Tukey–Kramer post-hoc test). In SHRs, a significantly greater number of PC shots was required after SEW than VPC treatment (1.5 fold, p < 0.05, Tukey–Kramer post-hoc test). c): NOS3 expression in all retinas of WWRs and SHRs before and after administration of each drug. The Y-axis indicate these are relative NOS3/gapdh to untreated control retina values. The expression of NOS3 was significantly increased in dimethyl sulfoxide (DMSO), SEW, and VPC -treated groups compared with the pre-treatment (control) group (p < 0.001, Tukey–Kramer post-hoc test). When comparing the expression of NOS3 between the SEW-treated WWRs and SHRs, the expression of NOS3 was significantly higher in the former (1.4 fold, p < 0.01, Tukey–Kramer post-hoc test).

Article Snippet: The WWRs and SHRs were each divided into four groups: control rats receiving no drug administration, rats treated with vehicle (dimethyl sulfoxide [DMSO]) only, rats treated with the S1PR1 agonist SEW2871 (SEW, Cayman Chemical, Ann Arbor, MI, USA), and rats treated with the S1PR1 antagonist VPC23019 (VPC, Cayman Chemical).

Techniques: Expressing, Quantitative RT-PCR, Control