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

Journal: Veterinary Medicine and Science

Article Title: FABP4 Mediates Inflammation to Regulate Endometrial Epithelial Cell Function During Early Gestation in Sheep

doi: 10.1002/vms3.70193

Figure Lengend Snippet: Primer sequences of target gene.

Article Snippet: The blots were incubated overnight at 4°C with the following primary antibodies: FABP4 rabbit polyclonal antibody (Proteintech, USA), Bcl2 rabbit polyclonal antibody (Proteintech, USA), NF‐κB rabbit polyclonal antibody (Proteintech, USA), β‐Actin mouse polyclonal antibody (Proteintech, USA), Phospho NF‐κB rabbit polyclonal antibody (Beyotime, China), Bax rabbit polyclonal antibody (Beyotime, China), and Caspase9 rabbit polyclonal antibody (Beyotime, China).

Techniques: Sequencing

Journal: Veterinary Medicine and Science

Article Title: FABP4 Mediates Inflammation to Regulate Endometrial Epithelial Cell Function During Early Gestation in Sheep

doi: 10.1002/vms3.70193

Figure Lengend Snippet: Culture and identification of endometrial epithelial cells isolated from sheep uteri. (A) The growth status of cells on the 2nd, 4th, and 6th days after tissue block inoculation. (B) Immunofluorescent identification of sEECs(CK18) (200×). The nuclei were stained with DAPI, and the epithelial cells were identified by observing the expression of CK18 (green fluorescence). (C) Identification of FABP4 protein expression in cells (200×). The nuclei were stained with DAPI, and FABP4 exhibited red fluorescence.

Article Snippet: The blots were incubated overnight at 4°C with the following primary antibodies: FABP4 rabbit polyclonal antibody (Proteintech, USA), Bcl2 rabbit polyclonal antibody (Proteintech, USA), NF‐κB rabbit polyclonal antibody (Proteintech, USA), β‐Actin mouse polyclonal antibody (Proteintech, USA), Phospho NF‐κB rabbit polyclonal antibody (Beyotime, China), Bax rabbit polyclonal antibody (Beyotime, China), and Caspase9 rabbit polyclonal antibody (Beyotime, China).

Techniques: Isolation, Blocking Assay, Staining, Expressing, Fluorescence

Journal: Veterinary Medicine and Science

Article Title: FABP4 Mediates Inflammation to Regulate Endometrial Epithelial Cell Function During Early Gestation in Sheep

doi: 10.1002/vms3.70193

Figure Lengend Snippet: The mRNA and protein expression levels of inflammatory factors in the control group, LPS group and LPS +BMS309403 group of sEECs hormonally treated to mimic the post‐implantation phase. (A–E) The mRNA expression levels of FABP4 , IL1β , IL6 , TNFα , and NF‐κB1 in the control group, LPS group, and LPS + BMS309403 group. (F–I) The protein expression levels of the FABP4 , P‐P65 , and P‐IκB proteins in the control group, LPS group, and LPS + BMS309403 group, as detected by western blot. All data are presented as mean ± standard error. Differences were considered significant at p < 0.05* and extremely significant at p < 0.01**.

Article Snippet: The blots were incubated overnight at 4°C with the following primary antibodies: FABP4 rabbit polyclonal antibody (Proteintech, USA), Bcl2 rabbit polyclonal antibody (Proteintech, USA), NF‐κB rabbit polyclonal antibody (Proteintech, USA), β‐Actin mouse polyclonal antibody (Proteintech, USA), Phospho NF‐κB rabbit polyclonal antibody (Beyotime, China), Bax rabbit polyclonal antibody (Beyotime, China), and Caspase9 rabbit polyclonal antibody (Beyotime, China).

Techniques: Expressing, Control, Western Blot

Journal: Veterinary Medicine and Science

Article Title: FABP4 Mediates Inflammation to Regulate Endometrial Epithelial Cell Function During Early Gestation in Sheep

doi: 10.1002/vms3.70193

Figure Lengend Snippet: Effects of FABP4‐controlled inflammation on the proliferation and migration of sEECs. (A) CCK8 was used to detect cell proliferation. (B–D) The mRNA expression levels of CCNA2 , CCNB1 , and CCND1 in the control group, the LPS group and the LPS + BMS309403 group were detected by RT‐qPCR. (E) Cell scratch assays were used to detect cell migration ability. (F–I) The mRNA expression levels of Vim , β‐cadherin, N‐cadherin , and E‐cadherin in the three groups were detected by RT‐qPCR. All data are presented as mean ± standard error. Differences were considered significant at p < 0.05* and extremely significant at p < 0.01**.

Article Snippet: The blots were incubated overnight at 4°C with the following primary antibodies: FABP4 rabbit polyclonal antibody (Proteintech, USA), Bcl2 rabbit polyclonal antibody (Proteintech, USA), NF‐κB rabbit polyclonal antibody (Proteintech, USA), β‐Actin mouse polyclonal antibody (Proteintech, USA), Phospho NF‐κB rabbit polyclonal antibody (Beyotime, China), Bax rabbit polyclonal antibody (Beyotime, China), and Caspase9 rabbit polyclonal antibody (Beyotime, China).

Techniques: Migration, Expressing, Control, Quantitative RT-PCR

Journal: iScience

Article Title: Vitamin D alleviates HFD-induced hepatic fibrosis by inhibiting DNMT1 to affect the TGFβ1/Smad3 pathway

doi: 10.1016/j.isci.2024.111262

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-FABP4 , Cell Signaling Technology , Cat#50699.

Techniques: Recombinant, Methylation, Enzyme-linked Immunosorbent Assay, cDNA Synthesis, Sequencing, Negative Control, Positive Control, Software