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Appendix S1. Laboratory methods for mitochondrial DNA.

Appendix S2. Nested structure analysis.

Appendix S3. Discussion of divergence time estimation assumptions.

Appendix S4. IM effective sample sizes and posterior probability distributions.

Appendix S5. Temporal analysis of the Saddle Road (1950 m) site.

Table S1. Nested analysis of populations using STRUCTURE 2.3.

Table S2. Cytochrome oxidase I (COI) rates of mutation.

Table S3. Effective sample sizes for each parameter from each independent IM run.

Table S4. Results of the clustering analysis that included individuals from 2005 to 2008 at Saddle Road (1950 m).

Table S5. Assignment to populations of individuals with mixed ancestry collected from Saddle Road (1950).

Figure S1. South Mauna Loa-Mauna Kea IM graphs: (A) m1 = migration rate into South Mauna Loa from Mauna Kea; (B) m2 = migration rate into Mauna Kea from South Mauna Loa; (C) t = maximum time of population splitting (unconverted).

Figure S2. South Mauna Loa-Mauna Kea IM graphs: (D) q1 = scalar for South Mauna Loa theta; (E) q2 = scalar for Mauna Kea theta; (F) qA = scalar for ancestral population theta.

Figure S3. Saddle Road (1950 m) High-Saddle Road Low (1950 m): (A) m1 = migration rate into Saddle Road (1950 m) High from Saddle Road Low (1950 m); (B) m2 = migration rate into Saddle Road Low (1950 m) from Saddle Road (1950 m) High; (C) t = maximum time of population splitting (unconverted).

Figure S4. Saddle Road (1950 m) High-Saddle Road Low (1950 m): (D) q1 = scalar for Saddle Road (1950 m) High theta; (E) q2 = scalar for Saddle Road Low (1950 m) theta; (F) qA = scalar for ancestral population theta.

Figure S5. Delta K values for the temporal clustering analysis.

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