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: Frontiers in Cell and Developmental Biology

Article Title: In vivo CRISPR-mediated activation of cardiogenic genes to reprogram cardiac fibroblasts

doi: 10.3389/fcell.2026.1812941

Figure Lengend Snippet: Activation of endogenous cardiogenic genes for fibroblast reprogramming. (A) Schematic of the CRISPRa system targeting the transcription start site of cardiogenic genes. The scFv (single-chain variable fragment) was engineered to recognize and bind the GCN4 peptide for transcriptional activation. (B,C) Quantitative PCR (B) and Western blot (C) analyses of cardiogenic gene expression in CFs isolated from tamoxifen-treated Pdgfra creERT2 ;Rosa-LSL-dCas9 ST transgenic mice after 7 days of transfection with AAVs carrying individual sgRNAs. Data are presented as mean ± standard error (n = 6 per group); Versus scramble sgRNA control: ns , not significant; * p < 0.05; ** p < 0.01. (D) Schematic illustration of the experimental design using CFs from Pdgfra creERT2 ;Rosa-LSL-dCas9 ST ;Nkx2.5CE eGFP mice, showing the induction procedure following tamoxifen treatment and infection with the AAV cocktail expressing sgRNAs. (E) Morphological changes in CRISPRa-induced and control CFs before and after suspension culture. Green fluorescence indicates activation of the Nkx2.5 CEeGFP reporter. Scale bars, 200 µm. (F) Representative immunostaining images for cardiac markers in CRISPRa GNTIS -induced cells after suspension culture and reattachment. Scale bars, 20 µm.

Article Snippet: Pdgfra CreER (Stock#032770), Rosa-LSL-dCas9 ST (Stock#031925), Nkx2.5CE eGFP (Stock#029489), and Rosa-LSL-tdTomato (Stock#007914) mice were obtained from The Jackson Laboratory.

Techniques: Activation Assay, Real-time Polymerase Chain Reaction, Western Blot, Gene Expression, Isolation, Transgenic Assay, Transfection, Control, Infection, Expressing, Suspension, Fluorescence, Immunostaining

Journal: Frontiers in Cell and Developmental Biology

Article Title: In vivo CRISPR-mediated activation of cardiogenic genes to reprogram cardiac fibroblasts

doi: 10.3389/fcell.2026.1812941

Figure Lengend Snippet: Assessment of cardiac function in MI mice. (A) Schematic of Pdgfra CreERT2 ;Rosa-LSL-dCas9 ST ;Rosa-LSL-tdTomato mice used to generate the MI model and for intramyocardial injection of AAVs expressing sgRNAs and scFv-VP64 during surgery. (B) Representative imaging of M-mode echocardiography at 8 weeks after surgery. (C–I) Quantification of cardiac function, including (C) left ventricular end-diastolic dimension (LVDd), (D) left ventricular end-systolic dimension (LVDs), (E) ejection fraction (EF), (F) fractional shortening (FS), (G) heart rate (HR), (H) stroke volume (SV), and (I) cardiac output (CO) measured at 2 or 8 weeks post-MI. Data are presented as mean ± standard error (n = 7 per group); ns , not significant; * p < 0.05.

Article Snippet: Pdgfra CreER (Stock#032770), Rosa-LSL-dCas9 ST (Stock#031925), Nkx2.5CE eGFP (Stock#029489), and Rosa-LSL-tdTomato (Stock#007914) mice were obtained from The Jackson Laboratory.

Techniques: Injection, Expressing, Imaging

Journal: Glia

Article Title: RXR Gamma Enables Oligodendrocyte Differentiation by Suppressing Sonic Hedgehog Signaling

doi: 10.1002/glia.70151

Figure Lengend Snippet: SHH signaling is dysregulated and differentiation is impaired in post‐natal Rxrg −/− OPC cultures. (A) Graph shows Shh gene expression in WT and Rxrg −/− primary OPC cultures. (B) Graph shows the percentage of SHH‐positive OPCs in WT and Rxrg −/− post‐natal primary OPC cultures. (C) Representative picture of WT and Rxrg −/− primary OPC (PDGFRa‐positive cells) undifferentiated cultures (no T3) double stained for SHH expression. Scale bar: 25 μm. (D) Graph shows the percentage of mature OLs in WT and Rxrg −/− post‐natal primary OPC cultures after exposure for 3 DIVs to the differentiation trigger T3. (E) Representative picture of WT and Rxrg −/− primary differentiated mature OLs (MBP‐positive cells). Scale bar: 25 μm. MBP, myelin basic protein; PDGFRa, platelet‐derived growth factor receptor alpha; T3, triiodothyronine; SHH, sonic hedgehog.

Article Snippet: Cells were resuspended in 80 μL of sorting buffer and 10 μL of blocking solution from PDGFRa Microbead kit (Miltenyi, Cat. N. 130‐101‐547).

Techniques: Gene Expression, Staining, Expressing, Derivative Assay

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Fibro‐Adipogenic Progenitors Regulate Orofacial Neuromuscular Junction Regeneration via Myostatin

doi: 10.1002/jcsm.70264

Figure Lengend Snippet: FAP‐specific MSTN knockout and pharmacological blockade of MSTN enhanced NMJ regeneration in MAS muscle. (a) AChR immunostaining of transverse sections from intact, PBS‐treated, or ACE‐031‐treated mice. Graphs in the right panel show the quantification of AChR counts per myofibre in MAS (upper) and TA (lower). n = 3 mice/group. (b) Masseter bite force in intact, PBS‐treated and ACE‐031‐treated mice at 30 dpi (upper). Hindlimb grip time assessed in intact, PBS‐treated and ACE‐031‐treated mice at 30 dpi (lower). n = 3 mice/group. (c) Schematic showing the generation of FAPs‐specific MSTN knockout mice (MSTN cKO). (d) AChR labelling of transverse sections of MAS or TA muscles of MSTN cKO mice (MSTN fl/fl ; Pdgfra Cre/+ ) and control mice (MSTN fl/fl ; Pdgfra +/+ ). Graphs in the right panel show the quantification of AChR counts per myofibre in MAS (upper) and TA (lower). n = 6 mice/group. (e) Masseter bite force (upper) and hindlimb grip time (lower) were measured in intact and injured MSTN cKO mice and controls. n = 3 mice/group. The data are shown as mean ± SD. One‐way ANOVA followed by Tukey's post hoc test was used. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: PDGFRa Cre/+ mice, MSTN fl/+ mice and ROSA nTnG mice (C57BL/6 background) were purchased from The Jackson Laboratory.

Techniques: Knock-Out, Immunostaining, Muscles, Control