rabbit monoclonal anti fabp4  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: TSPO deficiency reveals a novel role for porphyrins in regulating white adipose tissue lipid metabolism

doi: 10.1101/2025.09.14.676078

Figure Lengend Snippet: (A) Representative Oil Red O staining of primary adipocytes derived from Tspo fl/fl and Tspo -/- mice at Day 4 of diberentiation under adipogenic conditions indicated higher abundance of lipid accumulation in Tspo -/- adipocytes. (B) Graph showing quantification of lipid accumulation by Oil Red O absorbance at Day 0 and Day 4 of diberentiation. Tspo -/- adipocytes showed significantly higher levels of lipid accumulation compared to Tspo fl/fl controls. (C) Quantitative PCR analysis of Tspo mRNA expression during diberentiation (Days 0, 2, and 4) in Tspo fl/fl adipocytes demonstrates robust induction of Tspo transcript levels over the course of adipogenesis. (D) Western blot analysis of TSPO, PPARγ, and HSP90 protein levels at Days 0, 2, and 4 of diberentiation in Tspo fl/fl adipocytes indicated upregulation of TSPO and PPARγ at Day 2 and 4; HSP90 levels remained unchanged with diberentiation. Actin is shown as a loading control. (E) Western blot analysis of PPARγ and FABP4 protein expression in Day 4 adipocytes from two independent replicate samples of Tspo fl/fl and Tspo -/- mice. Tspo -/- cells show reduced PPARγ and FABP4 protein levels relative to controls. Actin serves as a loading control. (F) Relative mRNA expression of adipogenesis-associated genes ( Pparg, Fabp4, Cd36, Lpl, Glut4, Srebp1c, Fasn, Acaca, Hsl, Atgl ) in Tspo fl/fl and Tspo -/- adipocytes at Days 0 and 4. At Day 4, Tspo -/- adipocytes exhibit significant downregulation of Fabp4 , Acaca , Hsl , and Atgl , consistent with impaired adipogenic and lipolytic gene programs (mean ± SEM; *p < 0.05).

Article Snippet: Primary antibody incubations were performed overnight at 4°C using the following antibodies: rabbit anti-TSPO (EPR5384) monoclonal antibody (Abcam, #ab109497), rabbit anti-HSP90 (C45G5) monoclonal antibody (Cell Signaling, #4877), rabbit anti-PPARγ monoclonal antibody (Cell Signaling, #81B8), rabbit anti-FABP4 polyclonal antibody (Cell Signaling, #2120) and mouse monoclonal anti-β-actin antibody (Li-Cor, 926-42212) as a loading control.

Techniques: Staining, Derivative Assay, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Control

Journal: bioRxiv

Article Title: TSPO deficiency reveals a novel role for porphyrins in regulating white adipose tissue lipid metabolism

doi: 10.1101/2025.09.14.676078

Figure Lengend Snippet: (A) Baseline protoporphyrin IX (PPIX) concentrations measured in WAT was not diberent between Tspo fl/fl and Tspo -/- mice under standard chow conditions. (B) Relative mRNA expression of δ-aminolevulinic acid synthase ( Alas ) and ferrochelatase ( Fech ) in WAT under HF diet conditions showed significant downregulation in Tspo -/- mice compared to Tspo fl/fl controls. (C-D) During in vitro adipocyte diberentiation, Tspo -/- primary adipocytes showed significantly elevated expression of Alas and Fech at days 2 and 4 compared to Tspo fl/fl cells, suggesting dysregulation of porphyrin and heme biosynthetic genes in the absence of TSPO. (E) Adipogenic gene expression in Tspo fl/fl primary adipocytes treated with δ-aminolevulinic acid (ALA), protoporphyrin IX (PPIX), or hemin during diberentiation at Day 4. Treatment with ALA and PPIX significantly reduced mRNA levels of the adipogenic markers Pparg and Fabp4 , whereas hemin had no significant ebect. Data are presented as mean ± SEM. Diberent letters denote statistically significant diberences among groups (p < 0.05).

Article Snippet: Primary antibody incubations were performed overnight at 4°C using the following antibodies: rabbit anti-TSPO (EPR5384) monoclonal antibody (Abcam, #ab109497), rabbit anti-HSP90 (C45G5) monoclonal antibody (Cell Signaling, #4877), rabbit anti-PPARγ monoclonal antibody (Cell Signaling, #81B8), rabbit anti-FABP4 polyclonal antibody (Cell Signaling, #2120) and mouse monoclonal anti-β-actin antibody (Li-Cor, 926-42212) as a loading control.

Techniques: Expressing, In Vitro, Gene Expression