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. Author manuscript; available in PMC: 2023 May 1.
Published in final edited form as: Mol Carcinog. 2022 Feb 8;61(5):481–493. doi: 10.1002/mc.23392

Figure 6. Formation of xenograft by parental vs. NAT1 KO MDA-MB-231 cells following different engraftment protocols.

Figure 6.

NRGs mice were injected into the flank (panels A, D, and G) or the fat pads (panels B, E, and H) or via the tail vein (panels C, F, and I) with parental or NAT1 KO (KO2 and KO5) MDA-MB-231 breast cancer cells. Following flank and fat-pad injections, the primary tumors were allowed to grow to a pre-determined size prior to euthanasia. The mice which received tail-vein injections were euthanized after 20 days. A and B, the number of days taken to reach a pre-determined size (approximately 1 cm3 for flank injections and 0.75 cm3 for fat-pad injections). D and E, the weight of primary tumors at the time of euthanasia. G-I, the weight of lungs at the time of euthanasia. NAT1 KO cells had a slower growth rate in vivo, and as a result, it took more days to reach the pre-determined size, compared to the parental cells (panels A and B). At the time of euthanasia, the average weight of the primary tumors (panels D and E) or the lungs (panels G-I) did not differ significantly between the groups. Each data point represents an individual mouse and graphs illustrate mean ± SEM (n=5). Panels J-R, MDA-MB-231 parental and NAT1 KO cell lines were subcutaneous injected into the right flank and allowed to grow until the primary tumor had a size of about 1 cm3 (panels J, M and P). For the mammary fat-pad xenografts, the cells were injected subcutaneously under the right and left posterior nipples, and the primary tumors were allowed to grow until they reached approximate combined size of 0.75 cm3 (panels H, K, and N). For tail-vein injections animals were euthanized after 20 days (panels L, O, and R). Number of METs per lung (panels J-L), percent of the lung area containing METs (panels M-O), size of individual METs (panels P-R) in mice engrafted with the parental and NAT1 KO with MDA-MB-231 cells. Each data point represents an individual mouse and graphs illustrate mean ± SEM (n=5). *, significantly lower in mouse injected with NAT1 KO cells (p<0.05); **, p<0.01. P, parental; KO2, NAT1 KO cell line #2. KO5, NAT1 KO cell line #5.