(A) qPCR analysis of Slc39a8 expression in 12- to 14-week-old male Slc39a8^fl/fl and ZIP8-LSKO mice (n = 3–4). (B) ICP-OES analysis of Mn levels in 12- to 14-week-old male Slc39a8^fl/fl and ZIP8-LSKO mice (n = 4). (C) ICP-MS analysis of Mn levels in the whole blood of 14- to 16-week-old male Slc39a8^fl/fl and ZIP8-LSKO mice (n = 7 and 6, respectively). (D) ICP-OES analysis of Mn levels in 12- to 14-week-old male Slc39a8^fl/fl mice injected with AAV-null and in ZIP8-LSKO mice injected with AAV-null or AAV-ZIP8 (n = 3–6). (E) Western blot analysis of ZIP8 in liver lysates of male B6 mice injected with AAV-null or AAV-ZIP8 at 10 weeks of age and sacrificed 4 weeks after injection. Arrows indicate the ZIP8 bands. (F) ICP-OES analysis of Mn levels in male B6 mice injected with AAV-null or AAV-ZIP8 at 10 weeks of age and sacrificed 4 weeks after injection (n = 6). (G) ICP-MS analysis of Mn levels in the whole blood of male B6 mice injected with AAV-null or AAV-ZIP8 at 8 weeks of age and sacrificed 4 weeks after injection (n = 5 and 4, respectively). qPCR results were normalized to Gapdh. ICP-OES results were normalized to wet tissue weight. Mn levels were normalized to the average of the control group. Data on the absolute Mn content can be found in the Supplemental Table. (A–C, F, and G) Comparisons between 2 groups were performed using Student’s t test. Multiple comparisons in D were performed using 1-way ANOVA and Tukey’s multiple comparisons test. ***P ≤ 0.001, **P ≤ 0.01, and *P ≤ 0.05. (H) Correlation analysis between hepatic Mn and Mn levels in the kidney, brain, and heart of WT and ZIP8-LSKO mice and AAV-null– and AAV-ZIP8–injected B6 mice (n = 19). (I) Correlation analysis between hepatic Mn and Mn levels in the whole blood of WT and ZIP8-LSKO mice and AAV-null– and AAV-ZIP8–injected B6 mice (n = 22). Mn levels were normalized to the average of the control group. The results in H and I were analyzed by Pearson’s test. All data are shown as the mean ± SD.