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. 2022 Apr 15;12:6261. doi: 10.1038/s41598-022-10143-w

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Impaired osteolytic tumor growth of shGPR81-transduced MDA-MB-231 cells. (a) Soft X-ray analysis of the hind limbs of mice after bone marrow injections with 1 × 10⁵ shNT or shGPR81 #1 cells. The arrowheads indicate the osteolytic lesions. (b) Osteolytic destruction was quantified by measuring the areas of the osteolytic lesions in digital X-ray images. The data are presented as the percentage of the tibia occupied by osteolytic lesions (n = 9 mice/group). **p < 0.01 (vs. shNT cells); Student’s t-test. (c) Hematoxylin and eosin staining of histological sections of the tibiae collected from the mice shown in panel (a). B: cortical bone, *: tumor. Scale bars 500 μm (d) Tartrate-resistant acid phosphatase (TRAP) staining of serial sections presented in panel (c). Note that TRAP-positive osteoclasts aligned at the endosteal surfaces. B: cortical bone, *: tumor. Scale bars 500 μm (e) The number of TRAP-positive osteoclasts per bone surface (no. of TRAP + OC/BS) was measured using a light microscope (mean ± s.d., n = 5/group). **p < 0.01 (vs. shNT); Student’s t-test. (f) Total RNA was isolated from shNT and shGPR81 #1 cells, and IL6, IL11, and PTHrP mRNA expression was analyzed by RT-qPCR. Data are shown as fold changes normalized to shNT (mean ± s.d., n = 3/group). **p < 0.01 (vs. shNT); Student’s t-test.