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. 2022 Dec 21;13(12):1065. doi: 10.1038/s41419-022-05511-3

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — A, B Viability of 621-101 and 621-103 cells (upper panel) and ELT3-V3 and ELT3-T3 cells (lower panel) treated with the S1PR1 inhibitor W123 (A) or S1PR3 inhibitor TY52156 (B) at the indicated concentrations for 24 h was measured by MTT assay. (C) The death rate of 621-101 and 621-101 cells treated with TY52156 at the indicated concentrations for 24 h was measured by PI/CV assay. A–C Results are representative of eight independent samples per group. D The mRNA level of S1PR3 detected by RT-qPCR (upper panel) and the protein expression of S1PR3 and phosphor-S6 (235/236) detected by immunoblotting (lower panel) in vehicle control or TY52156-treated 621-101 cells were analyzed. E 621-101 cells treated with vehicle control or TY52156 (2 μM) were stained with Annexin V: FITC apoptosis detection kit reagent. Cell death was analyzed by flow cytometry (n = 3). F Migration (left panel) and invasion (right panel) of 621-101 cells treated with vehicle control or TY52156 (2 μM) were detected in Transwell chambers without or with Matrigel. G 621-101 cells were transfected with three independent S1PR3 siRNAs (#1, #2, or #3) or the negative control (siControl). The mRNA and protein levels of S1PR3 were determined using RT-qPCR (upper panel) and immunoblotting (lower panel). H, I Viability (H) and death rate (I) of 621-101siControl and 621-101siS1PR3 (#1, #2, or #3) cells were measured by MTT or PI/CV assay. The results are representative of eight independent samples per group. J 621-101siControl and 621-101siS1PR3 cells stably expressing the luciferase tag were injected intravenously into SCID mice (n = 5). Lung colonization was quantified using bioluminescence intensity 0 h, 6 h, and 24 h post injection. Luminescence color scale: 0 h, 6 h, and 24 h (min = 2 × 10⁵, max = 2 × 10⁶). The statistical analysis (right panel) indicates the relative fold change in photon flux. The results are representative of five mice per group. Student’s t test, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.