Figure EV3. Functional analysis of MitoT on CD3⁺ T cells. Metabolic changes, transmigration capacity and proliferation analysis on MitoT^pos cells.

• A–C
  Extracellular Acidification Rate (ECAR) analysis measured in a Seahorse XFp extracellular flux analyzer in FACS‐sorted MitoT^pos and MitoT^neg CD3⁺ T cells mitocepted with UC‐MSC mitochondria (n = 3 biological replicates, ran in quadruplicates).
• D–F
  Oxygen Consumption Rate (OCR) analysis measured in a Seahorse XFp extracellular flux analyzer in FACS‐sorted MitoT^pos and MitoT^neg CD3⁺ T cells mitocepted with UC‐MSC mitochondria (n = 3 biological replicates, ran in quadruplicates).
• G
  Glycolysis/OXPHOS ratio on FACS‐sorted MitoT^pos and MitoT^neg CD3⁺ T cells mitocepted with UC‐MSC‐MT (n = 3 biological replicates, ran in quadruplicates).
• H
  L(+)‐Lactate concentrations from supernatants of sorted CD3⁺ MitoT^neg and MitoT^pos T cells, previously mitocepted with UC‐MSC‐MT (n = 2 biological replicates, each one in triplicate).
• I
  Number of migrated CD4⁺ MitoT^neg and MitoT^pos sorted T cells from 100.000 cells seeded on media non‐activated or activated with anti‐CD3 and IL‐2 for 3 days (n = 3 biological replicates, in duplicates).
• J
  Percentage of CD3⁺ T cell proliferation after increasing Mitoception ratios, quantified by measuring the corresponding decrease in CellTrace Violet (CTV) proliferation dye intensity by flow cytometry (n = 3 biological replicates with three different PBMC donors).
• K
  Percentage of proliferation of FACS‐sorted MitoT^neg and MitoT^pos CD3⁺ cells, after 5 days in culture with anti‐CD3/CD28 beads (1/20) and IL‐2 (50 U/ml) in the presence or absence of PHA (15 μg/ml) (round or triangle dots respectively) (n = 5 biological replicates with five different PBMC donors).

Data information: All graphs show mean ± SEM and statistical analysis by Student's t‐test.