Figure 6. Hif1 β shRNA knockdown increases respiratory capacity during CoCl2 treatment.
A) Bioenergetics profile, as measured by oxygen consumption rate with a Seahorse X24 extracellular flux analyzer, of shHif1β-1, shHif1β-2, and shGFP 3T3-L1 adipocytes under normoxic conditions.
B) Bioenergetics profile, as measured by oxygen consumption rate with a Seahorse X24 extracellular flux analyzer, of shHif1β-1, shHif1β-2, and shGFP 3T3-L1 adipocytes after pre-treatment with CoCl2.
C) Basal respiration and maximal respiratory capacity of shHif1β-1, shHif1β-2, and shGFP 3T3-L1 adipocytes was determined by calculating the area under the curve (AUC) in the basal phase and after uncoupling with FCCP. Values are means ± SEM of 6-7 replicates of three separate experiments. Asterisks indicate a significant difference compared to shGFP control in all panels (p<0.05).
D) mRNA was isolated from shHif1β-1, shHif1β-2, and shGFP stably transfected adipocytes after eight days of differentiation, and subjected to normoxia, hypoxia (0.1% O2 for 16hrs), or 200 nM CoCl2 for 16 hrs. Expression of Cytc1 and Cox4.2 was measured by qPCR. Data shown as mean ± SEM of triplicate samples and repeated three times.