pparg  (Cell Signaling Technology Inc)

Journal: Bioactive Materials

Article Title: A foam cell-targeted lipophagy restoration strategy stabilizes vulnerable atherosclerotic plaques

doi: 10.1016/j.bioactmat.2026.02.041

Figure Lengend Snippet: Schematic of the anti-atherosclerotic mechanism of OPN-HMCN@MLT. ( A ) The study commenced with the synthesis of mesoporous carbon nanospheres (MCN) functionalized with an OPN-binding peptide and hyaluronic acid to construct the OPN-HMCN nanoplatform. The OPN-binding peptide was designed to recognize OPN enriched in the extracellular matrix and on the surface of foam cells, thereby enabling selective accumulation in OPN-rich pathological regions. Following OPN recognition, OPN-HMCN@MLT undergoes CD44-dependent endocytosis. Melatonin (MLT), a lipid autophagy–promoting agent, was subsequently encapsulated within the nanocarrier to form OPN-HMCN@MLT. Firstly, the released MLT can bind to and upregulate the expression of PPARα and PPARγ, which then promote the expression of downstream genes (ABCA1, ABCG1, ACOX-1, and CTP1A) and trigger the lipophagy. ( B ) Subsequently, its lipophagy-enhancing effects, including ABCA1/G1-mediated cholesterol efflux and CTP1A/ACOX-1-mediated mitochondrial fatty acid oxidation, were studied to confirm the reversal of foam cell formation. ( C ) These effects eventually promote foam cells to reverse into macrophages. Abbreviations: MCN, mesoporous carbon nanoparticle; OPN, osteopontin; MLT, melatonin; LDL, low-density lipoprotein; ox-LDL, oxidized low-density lipoprotein; PA, Photoacoustic.

Article Snippet: To block nonspecific binding, membranes were incubated with 5% skim milk for 1 h. Thereafter, membranes were incubated overnight at 4 °C with primary antibodies against ABCA1, ABCG1, ACOX1, CPT1A, LC3 (ab192890, 1:2000, abcam), LAMP1 (84658-5-RR, 1:8000, Proteintech), PPARα (66826-1-Ig, 1:3000, Proteintech), PPARγ (66936-1-Ig, 1:10000, Proteintech), P62 (18420-1-AP, 1:10000, Proteintech), MCAD (55210-1-AP, 1:3000, Proteintech), LCAD (17526-1-AP, 1:10000, Proteintech), tubulin (80762-1-RR, 1:10000, Proteintech), GAPDH (60004-1-Ig, 1:50000, Proteintech), and β-actin (66009-1-Ig, 1:20000, Proteintech).

Techniques: Binding Assay, Construct, Expressing

Journal: bioRxiv

Article Title: BAT protects against hepatic oxidative stress by remodeling the circulating metabolome

doi: 10.64898/2026.05.12.722834

Figure Lengend Snippet: A. Schematic overview of the strategy we used to study how BAT affects systemic metabolite and lipid profiles in vivo. Step 1, generating the BAT ablation mouse model ( Ucp1 -Cre; Pparg flox/flox , herein BAT-mice) and littermate control ( Pparg flox/flox , herein BAT+ mice); Step 2, collecting the serum samples from these mice under acute cold (6 °C for 1 and 3 h), chronic cold (6 °C for 3 days), and thermoneutrality (28 °C for 3 days), with ad libitum feeding; Step 3, quantifying the dynamic profiles of metabolites and lipids across different temperature conditions by LC-MS; Step 4, selecting the metabolites and lipids altered by BAT ablation and clustering them based on their profile similarity. The altered molecules included those accumulated in BAT-ablated mice, which might be cleared by BAT, and those elevated in mice with intact BAT, which might be derived from BAT. N = 8 for BAT-ablated mice and N = 10 for littermate controls. B-C. Correlation clustering of circulating metabolites ( B ) and lipids ( C ) altered by BAT ablation. An unpaired t-test was used to determine the significance of each circulating metabolite or lipid between BAT+ and BAT- mice under each temperature condition. A total of 206 metabolites and 249 lipids with p < 0.05 in at least one condition were included for correlation clustering. Clustering analysis was performed using Mfuzz package in R software, and pairwise Pearson correlation coefficients between molecules were used to generate heatmaps. Table S1 lists the p values and fold changes between the BAT+ and BAT- groups for these metabolites and lipids. N = 8 for BAT-ablated mice and N = 10 for littermate controls. BAT- > BAT+, molecules were accumulated in BAT-ablated mice. BAT+ > BAT-, molecules were elevated in littermate controls with intact BAT. BAT-, BAT-ablated mice; BAT+, littermate controls. C-1h, 6 °C for 1 hour; C-3h, 6 °C for 3 hours; C-3d, 6 °C for 3 days; TN-3d, 28 °C for 3 days. Mice were provided with ad libitum access to food and water throughout the experiment. AMP, adenosine monophosphate; γ-Glu-Glu, glutamyl-glutamic acid; Pyro-Glu, pyroglutamic acid, SAM, S-adenosylmethionine; GSSG, oxidized glutathione; GSH, reduced glutathione; GABA, γ-aminobutyric acid; TG, triglyceride; DG, diglyceride; PE, phosphatidylethanolamine; PC, phosphatidylcholine; SM, sphingomyelin; Cer, ceramide; EtherPE, ether-linked phosphatidylethanolamine; EtherPC, ether-linked phosphatidylcholine; CE, cholesterol ester; LPE, lysophosphatidylethanolamine; LPC, lysophosphatidylcholine; LNAPE, N-acyl-lysophosphatidylethanolamine; HexCer, hexosylceramide; CAR, acylcarnitine; OHFA, hydroxy fatty acid; DiOHFA, dihydroxy fatty acid; DHA, docosahexaenoic acid. D-E. Representative quantitative profiles of known BAT-regulated metabolites ( D ) and lipids ( E ). Relative units: quantitative peak areas from LC-MS normalized to the average. N = 8 for BAT-ablated mice and N = 10 for littermate controls. Statistic: unpaired t-test, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. Ad libitum conditions. C-1h, 6 °C for 1 hour; C-3h, 6 °C for 3 hours; C-3d, 6 °C for 3 days; TN-3d, 28 °C for 3 days. F. Representative quantitative profiles of metabolites and lipids newly identified as BAT-regulated in this study. Relative units: quantitative peak areas from LC-MS normalized to the average. N = 8 for BAT-ablated mice and N = 10 for littermate controls. Statistic: unpaired t-test, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. Ad libitum conditions. C-1h, 6 °C for 1 hour; C-3h, 6 °C for 3 hours; C-3d, 6 °C for 3 days; TN-3d, 28 °C for 3 days. G. Relative quantitative profiles of indicated fatty acids and 12, 13-diHOME in serum from BAT-ablated mice and littermate controls. Relative units: quantitative peak areas from LC-MS normalized to the average. N = 8 for BAT-ablated mice and N = 10 for littermate controls. Statistic: unpaired t-test. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. Ad libitum conditions. C-1h, 6 °C for 1 hour; C-3h, 6 °C for 3 hours; C-3d, 6 °C for 3 days; TN-3d, 28 °C for 3 days.

Article Snippet: PPARg flox/flox (Stock No. 004584), and Ucp1 -Cre (Stock No. 024670) mice were obtained from the Jackson Laboratory.

Techniques: In Vivo, Control, Liquid Chromatography with Mass Spectroscopy, Derivative Assay, Software

Journal: Advanced Science

Article Title: Caspase‐6 Controls Lipid and Energy Metabolism in Diet‐Induced Obesity

doi: 10.1002/advs.202514784

Figure Lengend Snippet: Caspase‐6 cleaves PPARγ and SP1 to control ATGL expression in adipocytes. (A, B) Immunoblot of PPARγ and ATGL proteins in eWAT (A) and iWAT (B) of WT and C6KO mice fed 60% HFD for 12 weeks (n = 4). Two tailed unpaired Student's t ‐test. (C) In vitro cleavage of recombinant PPARγ by active caspase‐6. Immunoblot of PPARγ. (D) Immunoblot of lysates from 3T3‐L1 adipocytes treated with cycloheximide (5 µg/ml) and TNFα (25 ng/ml) for indicated time with or without pretreatment of Emricasan (50 µg/ml) for 1 hr. (E, F) Immunoblot SP1 protein in eWAT (E) and iWAT (F) of WT and C6KO mice fed HFD for 12 weeks (n = 3). Two tailed unpaired Student's t ‐test. (G) In vitro cleavage of recombinant SP1 by active caspase‐6. Coomassie blue staining of gel. (H) Immunoblot of lysates from 3T3‐L1 adipocytes treated with cycloheximide (CHX, 5 µg/ml) and TNFα (25 ng/ml) for indicated time with or without pretreatment of Emricasan (50 µg/ml) for 1 hr. (I) Immunoblot of lysates from ex vivo cultured eWAT from WT or C6KO, treated with CHX (15 µg/ml) and TNFα (75 ng/ml) for indicated time. (J) Pnpla2 expression in 3T3‐L1 adipocytes treated with Rosiglitazone (5µM) or T0070907 (100 nM) in the presence or absence of Mithramycin (10 µM) for 6 h (n = 3). Two tailed unpaired Student's t ‐test. (K) In vitro cleavage of WT and mutant (D69E) PPARγ2 expressed in HEK293T by recombinant active caspase‐6. (L) In vitro cleavage of WT and mutant (D185E) SP1 expressed in HEK293T by recombinant active caspase‐6. (M) PPARγ activity reporter assay in HEK293T cell expressing PPRE‐H2B‐eGFP along with WT PPARγ or PPARγ D69E mutant in the absence or presence of CHX‐TNFα treatment (n = 8). Two tailed unpaired Student's t ‐test. (N‐V) Pearson correlation between CASP6 and PPARγ target genes in human adipose tissues ( GSE245948 ): Correlation matrix (N), AQP7 (O), CPT1B (P), FADS2 (Q), ME3 (R), PLIN4 (S), PCK2 (T), SLC27A1 (U), and SLC27A4 (V). (n = 76). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Recombinant active caspase‐6 was obtained from Enzo Life Sciences (#BML‐SE170), recombinant PPARγ from Sino Biological (#12019‐H20B), and recombinant SP1 from Active Motif (#81181).

Techniques: Control, Expressing, Western Blot, Two Tailed Test, In Vitro, Recombinant, Staining, Ex Vivo, Cell Culture, Mutagenesis, Activity Assay, Reporter Assay

Journal: Advanced Science

Article Title: Caspase‐6 Controls Lipid and Energy Metabolism in Diet‐Induced Obesity

doi: 10.1002/advs.202514784

Figure Lengend Snippet: Adipocyte‐specific caspase‐6 knockout protects against diet‐induced obesity and insulin resistance. Flox and adipocyte‐specific caspase‐6 knockout (ACKO) mice fed 60% HFD for 12 weeks. (A) Schematic diagram of generating Casp6 flox (Flox) mice and adipocyte‐specific Casp6 knockout (ACKO) mice. (B) Immunoblot of caspase‐6 protein in adipose tissues and livers. (C) Body weight (n = 9‐11). (D‐F) Tissue weights (n = 9‐11): eWAT (D), iWAT (E), BAT (F). Two tailed unpaired Student's t ‐test. (G–I) Indirect calorimetry (n = 6): oxygen consumption rate (G), carbon dioxide production (H), and energy expenditure (I). ANCOVA analysis with body weight as a covariate. (J, K) GTT (J, n = 9‐10) and ITT (K, n = 8‐9) with AUC quantification. GTT/ITT: two‐way ANOVA followed by Šídák's‐corrected post hoc test. (L) Immunoblots for the expression levels of PPARγ, SP1 and ATGL in eWAT and iWAT of Flox and ACKO mice fed HFD for 12 weeks. (M) Graphical summary. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Recombinant active caspase‐6 was obtained from Enzo Life Sciences (#BML‐SE170), recombinant PPARγ from Sino Biological (#12019‐H20B), and recombinant SP1 from Active Motif (#81181).

Techniques: Knock-Out, Western Blot, Two Tailed Test, Expressing