cell death activator  (MedChemExpress)

Journal: Journal of Lipid Research

Article Title: Aerobic glycolysis drives differentiation of unilocular adipocytes

doi: 10.1016/j.jlr.2026.101023

Figure Lengend Snippet: The HUVAS culture model facilitates the differentiation of unilocular human white adipocytes in vitro. A: Schematic of 2D, scaffold-free 3D, and scaffolded HUVAS cultures, done side by side using stromal vascular fraction cells from same donor and passage, cultured in the same media and with the same differentiation time, varying only the cell environment. B: Representative confocal images of BODIPY, CellMask, and DAPI-stained adipocytes on day (d)30. Scale bars for all images in Figure 1: 500 μm (main), 50 μm (inset). C–F: Quantification of lipid droplet diameter (C), total lipid droplet area (D), adipocyte unilocularity (E), and total adipocyte area (F) across isogenic 2D, 3D, and HUVAS cultures from n = 50 cells per spheroid and 3 spheroids per individual. Similar results were obtained from cells from three different cell donors. G, H: Capillary Western blot analysis on d30 of PLIN1, CIDEC, UCP1, PPARγ, FAS and ATGL protein and quantification normalized to total loaded protein (UCP1 blot shown here, all original blots shown in A,) from 3 technical replicates of the same individual. All total protein measurements (n = 3 per sample x 6 proteins of interest, n tot = 18) are shown in H (right panel). I: Representative confocal images of BODIPY, CellMask, and DAPI-stained 3D and HUVAS spheroids on d10, d20, d25 and d30. J, K: Quantification of lipid droplet diameter (J) and adipocyte unilocularity (K) across timepoints from n = 50 cells from one spheroid per condition, repeated at least two times. L: Relative mRNA expression levels of preadipocyte marker PDGFRA and adipogenesis marker CEBPA at d5, d10, d15, d20, d25 and d30 in HUVAS (left) and 3D (right) assessed by qPCR from 3 spheroids of the same individual and repeated twice with similar results. M: Relative mRNA expression of PPARG across timepoints in 3D compared to HUVAS analyzed as in L. N, O: Capillary Western blot for PPARγ and TOMM20 across timepoints, with quantification shown as raw chemiluminescence values by loading equal amounts of spheroid lysate, instead of equal protein amount, for each timepoint from 3 spheroids (technical replicates) of the same individual. P: mRNA expression levels of CIDEC and PLIN1 at d5, d10, d15, d20, d25 and d30 assessed by qPCR from >3 technical replicates of two different individuals. Mean and standard deviation (SD) shown in all applicable graphs throughout the paper unless specifically indicated otherwise.

Article Snippet: TaqMan primers for human PDGFRA (Hs00998018_m1), CEBPA (Hs00269972_s1), PPARG (Hs01115513_m1), PLIN1 (Hs00160173_m1), CIDEC (Hs01032998_m1) and B2M (Hs00187842_m1) were used.

Techniques: In Vitro, Cell Culture, Staining, Western Blot, Expressing, Marker, Standard Deviation

Journal: bioRxiv

Article Title: Sex- and hepatocyte PPARγ-dependent effects of an obesogenic dietary approach to induce MASH with fibrosis in mice

doi: 10.64898/2026.02.25.707976

Figure Lengend Snippet: Liver weight (A), liver TG content (B), liver cholesterol content (C), plasma ALT level (D), MASLD activity score (E), and picrosirius red-stained fibrotic area (F) of mice in cohorts #1 [LF, HF, and HFCF-fed mice] and #2 [LF, HF, and HFC+Fr-fed mice]. Representative pictures of hematoxylin & eosin (H&E)- and picrosirius red/fast green (SR/FG)-stained liver section of mice in cohort #1 (G, LF, HF, and HFCF-fed mice) and cohort #2 (H, LF, HF, and HFC+Fr-fed mice). Hepatic gene expression of Pparg (I), Cidec (J), Col1a1 (K), Timp1 (L), Tnfa (M), Ccl2 (N), Trem2 (O), Gnmt (P), Pemt (Q), and Bhmt (R) of mice in cohort #1 (HF, and HFCF-fed mice) and cohort #2 (HF, and HFC+Fr-fed mice). I-R, expression level is represented as relative values of HF-fed control mice in cohort #1 or coho rt #2. Data are shown as mean ± standard error of the mean (SEM). Letters (a-d) indicate significant differences between LF-fed and HF-fed control mice. Exclamation marks (!) indicate significant differences between HF-fed and HFCF-control mice in Cohort #1 or HF-fed and HFC+Fr-control mice in Cohort #2. Plus signs (+) indicate significant differences between HF-fed and HFCF- Pparg ΔHep mice in Cohort #1 or HF-fed and HFC+Fr- Pparg ΔHep mice in Cohort #2. Asterisks indicate significant differences between control and Pparg ΔHep mice. a, !, * p<0.05; b, !!, ++, ** p<0.01; !!!,+++, *** p<0.001; d, !!!!, ++++, ****, p<0.0001. n= 5-11 mice/group in cohort #1 and 7-8 mice/group in cohort #2.

Article Snippet: Then, ponceau S stain was removed, and the membranes were incubated in the blocking buffer: 5% non-fat dry milk in Tris-buffered saline with 0.1% Tween-20 (TBST) for one hour at room temperature, and incubated at 4°C overnight with either PPARγ (1:500, C26H12, Cell Signaling Technologies, Danvers, MA, USA), COL1A1 (1:1000, E8F4L, Cell Signaling Technologies,), CIDEC (1:500, NB100-430,Novus Biologicals, Centennial, CO), BHMT (1:10000, ab96415, Abcam, Waltham, MA).

Techniques: Clinical Proteomics, Activity Assay, Staining, Gene Expression, Expressing, Control

Journal: bioRxiv

Article Title: Sex- and hepatocyte PPARγ-dependent effects of an obesogenic dietary approach to induce MASH with fibrosis in mice

doi: 10.64898/2026.02.25.707976

Figure Lengend Snippet: Liver weight (A), liver TG content (B), liver cholesterol content (C), plasma ALT level (D), MASLD activity score (E), picrosirius red-stained fibrotic area of liver sections (F, SR), western blot for hepatic COL1A1 (G, top) with its Ponceau S staining (G, bottom) and quantification from male and female mice (G). Representative pictures of hematoxylin & eosin (H&E)- and picrosirius red (SR)-stained liver section of male (H) and female (I) mice. Hepatic gene expression of Pparg mRNA (J) and western blot for hepatic PPARγ (K, top) with its Ponceau S blot (K, bottom) and quantification of PPARγ1 and PPARγ2 from male and female mice (K). Hepatic gene expression of Cidea (L), Cidec (M), Ccl2 (N), Trem2 (O), Col1a1 (P), Timp1 (Q), Mat1a (R), Gnmt (S), Pemt (T), Ahcy (U), Bhmt (V), Cbs (W) of male and female. L-W, the expression level is represented as relative values of LF-fed control mice. Western blots for hepatic CIDEC (X, top) and BHMT (Y, top) with their Ponceau S staining (X, Y, bottom) and quantification from male and female mice. Data are shown as mean ± standard error of the mean (SEM). Letters (a-d) indicate significant differences between LF-fed and HFC+Fr-fed control mice. Asterisks indicate significant differences between HFC+Fr-fed control and HFC+Fr-fed Pparg ΔHep mice. a, * p<0.05; b, **, p<0.01; c,*** p<0.001; d, ****, p<0.0001. n= 4-9 male mice/group and 8-11 female mice/group.

Article Snippet: Then, ponceau S stain was removed, and the membranes were incubated in the blocking buffer: 5% non-fat dry milk in Tris-buffered saline with 0.1% Tween-20 (TBST) for one hour at room temperature, and incubated at 4°C overnight with either PPARγ (1:500, C26H12, Cell Signaling Technologies, Danvers, MA, USA), COL1A1 (1:1000, E8F4L, Cell Signaling Technologies,), CIDEC (1:500, NB100-430,Novus Biologicals, Centennial, CO), BHMT (1:10000, ab96415, Abcam, Waltham, MA).

Techniques: Clinical Proteomics, Activity Assay, Staining, Western Blot, Gene Expression, Expressing, Control

Journal: bioRxiv

Article Title: Sex- and hepatocyte PPARγ-dependent effects of an obesogenic dietary approach to induce MASH with fibrosis in mice

doi: 10.64898/2026.02.25.707976

Figure Lengend Snippet: Volcano plots showing the differentially expressed genes (DEG, padj<0.05) by HFC+Fr diet (A) and Pparg ΔHep (B) in male and female mice. Heatmaps showing the regulation of PPARγ-target genes: Cidea , Cidec , monoacylglycerol O-acyltransferase 1 ( Mogat1 ), fatty acid translocase ( Cd36 ), perilipin 4 ( Plin4 ), the inflammation-related genes: Tnfa , Ccl2 , Trem2 , the fibrosis-related genes: Col1a1 , metalloproteinase 12 ( Mmp12 ), Timp1 , and the methionine metabolism-related genes: Pemt , Gnmt , Bhmt , by HFC+Fr diet (C) and Pparg ΔHep (D) in male and female mice. Asterisk (p<0.05) indicate significant regulation. Selected groups of genes identified by the enrichment analysis of DEG by HFC+Fr diet (E) and Pparg ΔHep (F) in male and female mice. The number of upregulated (green) or downregulated (red) DEG in the volcano plots or enrichment analysis are indicated in brackets. n= 4-6 male mice/group and 5 female mice/group.

Article Snippet: Then, ponceau S stain was removed, and the membranes were incubated in the blocking buffer: 5% non-fat dry milk in Tris-buffered saline with 0.1% Tween-20 (TBST) for one hour at room temperature, and incubated at 4°C overnight with either PPARγ (1:500, C26H12, Cell Signaling Technologies, Danvers, MA, USA), COL1A1 (1:1000, E8F4L, Cell Signaling Technologies,), CIDEC (1:500, NB100-430,Novus Biologicals, Centennial, CO), BHMT (1:10000, ab96415, Abcam, Waltham, MA).

Techniques:

Journal: medRxiv

Article Title: A dominant CIDEC variant segregates with familial obesity by increasing lipid droplet size in adipocytes

doi: 10.1101/2025.07.28.25332029

Figure Lengend Snippet: Panel A shows the pedigree of a family with progressive fat accumulation across generations. Squares indicate males, circles females, open symbols unaffected, black symbols affected and diagonal slash deceased. After whole exome sequencing, the mutant gene was identified: ‘+’ denotes wildtype, and ‘mut.’ for mutant alleles. Panel B shows images of the back, upper arm and thighs from affected individuals including the proband (I:1), his daughter (II:4) and a granddaughter (III:1). Panel C depicts a puncture biopsy taken from the I:1 proband’s thigh, revealing the unilocular adipocytes structures in the affected tissues. Panel D shows X-ray images of the proband’s daughter II:4, highlighting the accumulation of fat subcutaneously. Panel E shows a schematic of the germline mutation c.37A>G; p.(Arg13Gly) missense variant specific to the long isoform of CIDEC. Panel F depicts the strict phylogenetic conservation of the CIDEC-L R13 residue across evolution. Panel G shows the Minor allele frequency (MAF) against the combined annotation-dependent depletion (CADD) score for coding variants found in the first 13 residues of CIDEC-L as listed in gnomAD v.4.0.0 (black and grey dots) and CIDEC-L R13G found in our family (orange). Notably CIDEC-L R13G clusters with loss of function variants.

Article Snippet: Cidec point mutation knock-in mice were generated on a C57Bl/6J genetic background by Cyagen Biosciences (USA) and kept on a C57Bl/6J genetic background.

Techniques: Sequencing, Mutagenesis, Variant Assay, Residue

Journal: medRxiv

Article Title: A dominant CIDEC variant segregates with familial obesity by increasing lipid droplet size in adipocytes

doi: 10.1101/2025.07.28.25332029

Figure Lengend Snippet: Panel A shows the overexpression of GFP-tagged CIDEC variants in 3T3-L1 cells. Representative immunofluorescence images are depicted on the left, with LipidTox stained LDs (red). Introducing the p.R13G patient mutation into CIDEC-L causes an increase of LD size as quantified on the right (Kruskal-Wallis Test). This can likely be attributed to an increase of the lipid exchange rate (under co-transfection with PLIN1 , unpaired two-tailed t-test, Panel B ) and an increased number of lipid droplet contact sites per cell (LDCS, Kruskal-Wallis Test, Panel C ). The localization of CIDEC-L to LDCS is not affected by the p.R13G variant (one-way ANOVA, p>0.05, Panel D ). * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001 for all tests.

Article Snippet: Cidec point mutation knock-in mice were generated on a C57Bl/6J genetic background by Cyagen Biosciences (USA) and kept on a C57Bl/6J genetic background.

Techniques: Over Expression, Immunofluorescence, Staining, Mutagenesis, Cotransfection, Two Tailed Test, Variant Assay

Journal: medRxiv

Article Title: A dominant CIDEC variant segregates with familial obesity by increasing lipid droplet size in adipocytes

doi: 10.1101/2025.07.28.25332029

Figure Lengend Snippet: We performed co-immunoprecipitation (Co-IP) of N-terminal CIDEC variants to demonstrate that CIDEC-S and CIDEC-L form homo-mers ( Panel A ). The homotypic binding of CIDEC-L is not impaired by the R13G mutation (lane 4). Moreover, CIDEC-L and CIDEC-L R13G retain the ability to bind to CIDEC-S and CIDEA as shown by Co-IP in panel B .

Article Snippet: Cidec point mutation knock-in mice were generated on a C57Bl/6J genetic background by Cyagen Biosciences (USA) and kept on a C57Bl/6J genetic background.

Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Binding Assay, Mutagenesis

Journal: medRxiv

Article Title: A dominant CIDEC variant segregates with familial obesity by increasing lipid droplet size in adipocytes

doi: 10.1101/2025.07.28.25332029

Figure Lengend Snippet: We co-expressed fluorescently tagged CIDEC isoforms in 3T3-L1 cells, showing that the p.R13G mutation does not impact CIDEC-L’s ability to co-localize with CIDEC-S ( Panel A ). LDs size quantifications are shown on the right. Importantly, CIDEC-L and CIDEC-L R13G , but not CIDEC-S, also co-localize with CIDEA when overexpressed in 3T3-L1 cells ( Panel B ). LDs size quantifications are also on the right.

Article Snippet: Cidec point mutation knock-in mice were generated on a C57Bl/6J genetic background by Cyagen Biosciences (USA) and kept on a C57Bl/6J genetic background.

Techniques: Mutagenesis

Journal: medRxiv

Article Title: A dominant CIDEC variant segregates with familial obesity by increasing lipid droplet size in adipocytes

doi: 10.1101/2025.07.28.25332029

Figure Lengend Snippet: Panel A shows the schematic of the C57BL/6J mouse model with point mutation (c.28A>G, AGG to GGG, red highlight) at mouse Cidec locus by CRISPR/Cas9-mediated genome engineering. Synonymous mutation (highlighted in Magenta) was introduced to prevent the binding and re-cutting of the sequence by gRNA after homology-directed repair. Like the human mutation, the mouse c.28A>G point mutation selectively alters the Cidec-L protein sequence (p.R10G), without affecting the Cidec-S protein. Panel B shows gDNA sequences confirmed by Sanger sequencing. Panel C shows mRNA expression of Cidec-S , Cidec-L , Ucp1 and Cidea gene expression in tissues of iBAT, iWAT, gWAT and liver from 6 weeks old wildtype mice housed at 23 °C. *p < 0.05, **p <0.01, ****p < 0.0001 were calculated using one-way ANOVA with Dunnett’s multiple comparisons test (n = 5/group). Data are shown as individual measurements and results are presented as means ± SEM. Panel D shows immunoblot of endogenous Cidec protein expression in tissues of iBAT and iWAT of 6 weeks old wildtype mice housed at 23 °C, n = 2 mice for each tissue.

Article Snippet: Cidec point mutation knock-in mice were generated on a C57Bl/6J genetic background by Cyagen Biosciences (USA) and kept on a C57Bl/6J genetic background.

Techniques: Mutagenesis, CRISPR, Binding Assay, Sequencing, Expressing, Gene Expression, Western Blot