Figure 1|. Lasmiditan Enhances Mitochondrial Maximal Respiration Rate & Number in Primary MRPECs.

(A) PCR assessment of primary MRPEC monocultures for the expression of HTR1F. (B) Seahorse XF96 analyses showing the percent increase in MRPECs maximal mitochondrial respiration rate after FCCP [5 uM] injection, after 24 hrs of lasmiditan treatment [100 nM]. Data are representative of the average percent increase (± SEM) from basal OCR readings following FCCP [5 uM] injection across N=6 MRPEC replicates/mice and N=6 mice/treatment condition. RT-qPCR analyses of MRPECs after 24 hrs of lasmiditan treatment [100 nM] were then employed to determine relative mRNA expression changes across various mitochondrial biogenesis related genes: (C) PGC-1α, (D) TFAM, and (E) ATP Synthase-β. (F) Representative images of MRPEC monocultures after 24 hrs of lasmiditan [100nM] or vehicle treatment. Mitochondria are shown stained with MitoTracker Red CMXRos, binary processed, and then skeletonized to produce a skeleton tree for downstream Mitochondrial Analyzer Plug-in Analyses in ImageJ: (G) mitochondrial number/field, (H) mitochondrial area/field, (I) mitochondrial perimeter/field, and (J) mitochondrial mean form factor/mitochondria/field. One-Way ANOVA followed by a Tukey post-hoc correction for multiple comparisons was employed to identify significant FCCP-OCR alterations across various lasmiditan concentrations. All other data within this figure underwent Welch’s two-sample t-tests to identify significant differences in mRNA expression and mitochondrial features between treatment groups. A p-value of p≤0.05 was considered significant between treatment groups and is denoted by asterisks (*). Data are representative of the average fold change from vehicle-treated control groups (± SEM) across N=6 MRPEC isolations/treatment group where N=6 mice/treatment condition.