Figure 1.
Figure 1.

Heat stress (HS) stimulates adipogenesis in adipocytes. (A) oil red O staining of pig adipocytes on d 3, 6, and 9 of differentiation (40x) at 37 and 41.5°C temperatures. (B) Triglyceride (TAG) concentration was measured in pig adipocytes under normal conditions (37°C) and HS condition (41.5°C) on d 3, 6, and 9, respectively. Bars represent means ± SEM of at least 6 different replicates. *Mean difference between HS and control at P < 0.05.

 


Figure 2.
Figure 2.

Expression of genes related to adipogenesis in differentiating adipocytes under normal (37°C) and heat stress (41.5°C) conditions. (A) Peroxisome proliferator-activated receptor γ (PPARγ) gene expression. (B) CCAAT/enhancer binding proteins α (C/EBPα) gene expression. (C) PPARγ immunoblotting (upper panel) and quantification (lower panel). Bars represent means ± SEM of at least 6 different replicates. *Mean difference between heat stress and control at P < 0.05.

 


Figure 3.
Figure 3.

Expression of genes involved in fatty acid transport and binding and glucose transport in adipocytes under normal condition (37°C) and heat stress condition (41.5°C) on d 3, 6, and 9, respectively. Genes were measured by real-time PCR. (A) Fatty acid synthase (FAS). (B) Adipocyte protein 2 (aP2). (C) Fatty acid translocase 36 (CD36). (D) Lipoprotein lipase (LPL). (E) Fatty acid transport protein 4 (FATP4). (F) Fatty acid transport protein 6 (FATP6). (G) Glucose transporter protein type 4 (GLUT4). Bars represent means ± SEM of at least 6 different replicates. *Mean difference between heat stress and control at P < 0.05.

 


Figure 4.
Figure 4.

Heat stress increases abundance of genes involved in glyceroneogenesis in adipocytes. (A) Phosphoenolpyruvate carboxykinase 1 (PCK1 or PEPCK-C) and (B) glycerol kinase (GK) mRNA levels were measured by real-time PCR. (C) and (D) are immunoblots for PCK1 and GK, respectively. Bars represent means ± SEM of at least 6 different replicates. *Mean difference between heat stress and control at P < 0.05.

 


Figure 5.
Figure 5.

Heat stress stimulates adipokines expression in pig adipocytes. (A) Leptin and (B) adiponectin mRNA abundance was measured by real-time PCR in adipocytes under normal (37°C) and heat stress (41.5°C) conditions. (C) Media leptin was measured by ELISA. (D) Media adiponectin was measured by western blotting with quantification. Bars represent means ± SEM of at least 6 different replicates. *Mean difference between heat stress and control at P < 0.05.

 


Figure 6.
Figure 6.

Heat stress stimulates heat shock protein (HSP) expressions in pig adipocytes. Transcript levels of HSP 27(A), HSP60 (B), HSP70 (C) and HSP90 (D) were measured in pig adipocytes under normal (37°C) and heat stress (41.5°C) conditions using real-times PCR. (E) HSP70 and (F) HSP90 protein expression was determined by western blot. Bars represent means ± SEM of at least 6 different replicates. *Mean difference between heat stress and control at P < 0.05.

 


Figure 7.
Figure 7.

Heat stress leads to increase cellular ATP concentration. Adenosine triphosphate level was measured in pig adipocytes under normal (37°C) and heat stress (41.5°C) conditions. Bars represent means ± SEM of at least 6 different replicates. *Mean difference between heat stress and control at P < 0.05.