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. 2018 Aug 8;33(2):469–486. doi: 10.1038/s41375-018-0222-x

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© The Author(s) 2018

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. 3 — TRIM44 expression increases quiescent MM cell engraftment in the OS niche. a TRIM44^OE-CON and TRIM44^OE MM cells (RPMI, U266) were labeled with PKH26 and were i.v. injected into irradiated NOD/SCID mice. After 60 h, BM cells were separated into the OS or VS niches, and PKH⁺ MM cell numbers were calculated using FACS. Total 12 mice were used in this experiment. Detailed mice numbers in each experimental group are described in Table S7. b TRIM44^KD-CON and TRIM44^KD MM cells (RPMI, U266) were labeled with PKH26, and were i.v. injected into irradiated NOD/SCID mice. After 60 h, BM cells were separated into the OS or VS niches, and PKH⁺MM cell numbers were calculated using FACS. Total 12 mice were used in this experiment. Detailed mice numbers in each experimental group are described in Table S8. c NOD/SCID mice were i.p. injected with PTH (50 μg/kg) or vehicle for 21 days. PKH26 labeled TRIM44^OE–CON and TRIM44^OE MM cells (RPMI and U266) were then i.v. injected into the mice. After 60 h, BM cells were isolated as the OS and VS niche followed by FACS analyses. Total 24 mice were used in this experiment. Detailed mice numbers in each experimental group are described in Table S9. d NOD/SCID mice were i.p. injected with PTH or vehicle for 21 days. PKH26 labeled TRIM44^KD-CON and TRIM44^KD MM cells (RPMI and U266) were then i.v. injected into the mice. After 60 h, BM cells were isolated as the OS and VS nice followed by FACS analyses. Total 24 mice were used in this experiment. Detailed mice numbers in each experimental group are described in Table S9. e The recovery rates of PKH26⁺ TRIM44^OE-CON or TRIM44^OE MM cells in both the OS and VS niches were calculated in AMD3100 pre-treated mice. NOD/SCID mice were treated i.p. with AMD3100 for 5 days, followed by an i.v. injection with PKH26 labeled TRIM44^OE-CON or TRIM44^OE cells (1 × 10⁶). After engraftment for 60 h, MM cell recovery rates were calculated based on PKH⁺ staining using FACS. The statistical significance was calculated using an unpaired Student’s t-test and is represented as a P-value. Total 12 mice were used in this experiment. Detailed mice numbers in each experimental group are described in Table S10