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. 2023 Oct 11;12:e86452. doi: 10.7554/eLife.86452

Figure 7. GDF15 is required to regulate core body temperature in cold-exposed OPA1 BKO mice.

(A) Averaged core body temperature (light and dark cycles) collected from 12-week-old wild-type (WT) and OPA1/GDF15 brown adipose tissue (BAT) double-knockout (DKO) mice after 7 d at 30°C. (B) Averaged core body temperature (light and dark cycles) in mice cold exposed for 24 hr (4°C). (C) Final core body temperature recorded by telemetry in mice exposed to 4°C in the CLAMS system. (D–G) Indirect calorimetry data represented as the average for the light and dark cycles during the first 24 hr of data recording (4°C). (D) Energy expenditure. (E) Respiratory exchange ratio. (F) Food intake. (G) Locomotor activity. (H) Hourly core body temperatures collected from 12-week-old WT and DKO mice during cold exposure (4°C). (I–N) Data collected after 5 hr of cold exposure. (I) Relative mRNA expression of thermogenic genes in BAT after 5 hr of cold exposure normalized to tata box protein (Tbp). (J) Representative immunoblots for UCP1 in BAT normalized to β-actin and their respective densitometric quantification. (K) Pyruvate-malate-supported oxygen consumption rates (OCRs) and UCP1-dependent respirations in mitochondria isolated from BAT (baseline conditions). (L) Relative mRNA expression of thermogenic genes in inguinal white adipose tissue (iWAT) normalized to Tbp expression. (M) Representative immunoblots for UCP1 in iWAT normalized to β-actin and their respective densitometric quantification. (N) Representative immunoblots for Serca1a in gastrocnemius muscle normalized to Ponceau red staining and their respective densitometric quantification. Optical density (OD). Data are expressed as means ± SEM. Significant differences were determined by Student’s t-test using a significance level of p<0.05. * p <0.05; ** p <0.01. Significantly different vs. WT mice.

Figure 7—source data 1. GDF15 is required to regulate core body temperature in cold-exposed OPA1 BKO mice (uncropped blots with the relevant bands labeled).
(G) Full immunoblot images for UCP1 and β-actin in brown adipose tissue (BAT). (M) Full immunoblot images for UCP1 and β-actin in inguinal white adipose tissue (iWAT). (N) Full immunoblot for Serca1a and β-actin in gastrocnemius muscle.
Figure 7—source data 2. Original file with the full raw unedited blot for UCP1 in brown adipose tissue (BAT) of OPA1/GDF15 double-knockout (DKO) mice after cold exposure.
Figure 7—source data 3. Original file with the full raw unedited blot for β-actin for UCP1 in brown adipose tissue (BAT) of OPA1/GDF15 double-knockout (DKO) mice after cold exposure.
Figure 7—source data 4. Original file with the full raw unedited blot for UCP1 in inguinal white adipose tissue (iWAT) of OPA1/GDF15 double-knockout (DKO) mice after cold exposure.
Figure 7—source data 5. Original file with the full raw unedited blot for β-actin for UCP1 in inguinal white adipose tissue (iWAT) of OPA1/GDF15 double-knockout (DKO) mice after cold exposure.
Figure 7—source data 6. Original file with the full raw unedited blot for Serca1 in gastrocnemius muscle of OPA1/GDF15 double-knockout (DKO) mice after cold exposure.
Figure 7—source data 7. Original file with the full raw unedited blot for β-actin for Serca1a in gastrocnemius muscle of OPA1/GDF15 double-knockout (DKO) mice after cold exposure.

Figure 7.

Figure 7—figure supplement 1. Indirect calorimetry, food intake, locomotor activity, and skeletal muscle characterization of cold-exposed OPA1/GDF15 brown adipose tissue (BAT) double-knockout (DKO) and OPA1 BAT knockout (KO) mice.

Figure 7—figure supplement 1.

(A–H) Hourly indirect calorimetry data collected in OPA1 BKO and OPA1/GDF15 BAT DKO mice and their respective wild-type (WT) controls during the first 24 hr of cold exposure. (A) Oxygen consumption in OPA1 BKO mice. (B) Oxygen consumption in DKO mice. (C) Respiratory exchange ratio in OPA1 BKO mice. (D) Respiratory exchange ratio in DKO mice. (E) Food consumed in OPA1 BKO mice. (F) Food consumed in DKO mice. (G) Locomotor activity in OPA1 BKO mice. (H) Locomotor activity in DKO mice. (I) Representative immunoblots for tyrosine hydroxylase (TH) in inguinal white adipose tissue (iWAT) from mice exposed to 4°C for 5 hr normalized to β-actin and their respective densitometric quantification. (J) Representative immunoblots for GLUT1 in skeletal muscle from mice exposed to 4°C for 5 hr normalized to β-actin and their respective densitometric quantification. Optical density (OD). Data are expressed as means ± SEM. Significant differences were determined by ANCOVA for the group effect.
Figure 7—figure supplement 1—source data 1. I Full immunoblot images for tyrosine hydroxylase (TH) and β-actin in inguinal white adipose tissue (iWAT) of wild-type (WT) and OPA1/GDF15 brown adipose tissue (BAT) double-knockout (DKO) mice.
(J) Full immunoblot images for GLUT1 and β-actin in skeletal muscle of WT and OPA1/GDF15 BAT DKO mice.
Figure 7—figure supplement 1—source data 2. Original immunoblot image for tyrosine hydroxylase in inguinal white adipose tissue (iWAT) of wild-type (WT) and OPA1/GDF15 brown adipose tissue (BAT) double-knockout (DKO) mice.
Figure 7—figure supplement 1—source data 3. Original immunoblot image for β-actin for tyrosine hydroxylase in inguinal white adipose tissue (iWAT) of wild-type (WT) and OPA1/GDF15 brown adipose tissue (BAT) double-knockout (DKO) mice.
Figure 7—figure supplement 1—source data 4. Original immunoblot image for GLUT1 in gastrocnemius muscle of wild-type (WT) and OPA1/GDF15 brown adipose tissue (BAT) double-knockout (DKO) mice.
Figure 7—figure supplement 1—source data 5. Original immunoblot image for β-actin for GLUT1 in gastrocnemius muscle of wild-type (WT) and OPA1/GDF15 brown adipose tissue (BAT) double-knockout (DKO) mice.