Journal: bioRxiv

Article Title: Lineage Tracing Reveals a Shared Cellular Origin for Supraclavicular Brown and Inguinal Beige Adipocytes

doi: 10.1101/2025.10.02.680118

Figure Lengend Snippet: (A) An experimental design for isolating Mef2c-AHF + cells from scBAT of postnatal Mef2c-AHF- Cre;ROSA mTmG mice. mG + : Mef2c-AHF + cells. mT + : tdTomato + cells. (B) Representative FACS plots of SVF isolated from the scBAT of postnatal 2.5- to 5-week-old Mef2c-AHF-Cre;ROSA mTmG mice. n = 28. (C) A schematic representation of the brown adipocyte differentiation procedure. Rosi: rosiglitazone. Iso: isoproterenol. (D) Representative images of freshly sorted and spontaneously differentiated Mef2c-AHF + cells. The presence of lipid droplets in these cells is indicated by Oil-Red-O or LipidSpot 610. D8: day 8. Scale bar = 10 μm. n = 3-4. (E) Representative images of Mef2c-AHF + cells treated with IM or Bmp7+Rosi. The presence of lipid droplets in these cells is indicated by Oil-Red-O or LipidSpot 610. Scale bar = 10 μm. n = 3-5. (F) Relative mRNA levels of genes involved in brown adipocyte differentiation and mitochondrial biogenesis in Mef2c-AHF + cells that differentiated spontaneously or were induced by IM or Bmp7+Rosi. Iso was added to stimulate thermogenesis at the end of the culture period. Data are presented as mean ± SEM. **, p < 0.01; *, p < 0.05; ns = nonsignificant. n = 3-4.

Article Snippet: The antibodies used for western blotting included PPARγ (Cell Signaling, #2430S, 1:1000 or Santa Cruz, #sc-7273, 1:500), FABP4 (Abcam, 13979, 1:2000), UCP1 (Abcam, #10983, 1:2000), BMP7 (Cell Signaling, #4693S, 1:1000), BMPR1A (Proteintech, #12701-1-AP, 1:1000), BMPR2 (Cell Signaling, #6979S, 1:1000), SMAD1 (Cell Signaling, #9743S, 1:1000) and β-ACTIN (Cell Signaling, #4976S, 1:2000), and HRP (Jackson ImmunoResearch, Code#715-035-150, 1:3000 or Code#715-035-152, 1:3000).

Techniques: Isolation

Journal: bioRxiv

Article Title: Lineage Tracing Reveals a Shared Cellular Origin for Supraclavicular Brown and Inguinal Beige Adipocytes

doi: 10.1101/2025.10.02.680118

Figure Lengend Snippet: (A) A schematic diagram of the FACS strategy for isolating CD34 - cells from scBAT of the postnatal wild-type mice. (B) Representative density plots showing FACS staining profiles and gating (black boxes) of SVF isolated from scBAT. n = 40. (C) Representative images of Oil-Red-O-stained CD34 - cells that either differentiated spontaneously or were induced by IM or by Bmp7+Rosi. n = 3. (D) Representative fluorescent images of BODIPY493/503-stained CD34 - cells that either differentiated spontaneously or were induced by IM or by Bmp7+Rosi. Scale bar = 10 μm. n = 3-4. (E) Relative mRNA levels of genes involved in brown adipocyte differentiation and mitochondrial biogenesis in CD34 - cells that differentiated spontaneously or were induced by IM or by Bmp7+Rosi. Iso was added to stimulate thermogenesis at the end of the culture period. Data are presented as mean ± SEM. ***, p < 0.001; **, p < 0.01; *, p < 0.05; ns = nonsignificant. n = 3-5.

Article Snippet: The antibodies used for western blotting included PPARγ (Cell Signaling, #2430S, 1:1000 or Santa Cruz, #sc-7273, 1:500), FABP4 (Abcam, 13979, 1:2000), UCP1 (Abcam, #10983, 1:2000), BMP7 (Cell Signaling, #4693S, 1:1000), BMPR1A (Proteintech, #12701-1-AP, 1:1000), BMPR2 (Cell Signaling, #6979S, 1:1000), SMAD1 (Cell Signaling, #9743S, 1:1000) and β-ACTIN (Cell Signaling, #4976S, 1:2000), and HRP (Jackson ImmunoResearch, Code#715-035-150, 1:3000 or Code#715-035-152, 1:3000).

Techniques: Staining, Isolation

Journal: bioRxiv

Article Title: Lineage Tracing Reveals a Shared Cellular Origin for Supraclavicular Brown and Inguinal Beige Adipocytes

doi: 10.1101/2025.10.02.680118

Figure Lengend Snippet: (A) Representative images of X-gal-stained hearts from E12.5 Mef2c-AHF-Cre;R26R and Prrx1-Cre;R26R embryos. Scale bar = 2000 μm. n = 4. ra: right atrium, rv: right ventricle, oft: outflow tract, la: left atrium, lv: left ventricle. (B) Representative fluorescence images of scBAT and iBAT from E18.5 and postnatal Prrx1-Cre;ROSA mTmG mice. Scale bar = 25 μm. n = 3-4. (C) Experimental design for isolating Prrx1 + cells from the scBAT of postnatal Prrx1-Cre;ROSA mTmG mice. mG + : Prrx1 + cells. mT + : tdTomato + cells. (D) Representative FACS plots of SVF isolated from scBAT of the postnatal Prrx1-Cre;ROSA mTmG mice. n = 20. (E) Representative images of Oil-Red-O and LipidSpot 610-stained Prrx1 + cells that differentiated spontaneously or were induced by IM or by Bmp7+Rosi. Scale bar: 10 μm. n = 4-5. (F) Relative mRNA levels of genes involved in brown adipocyte differentiation and mitochondrial biogenesis in Prrx1 + cells that differentiated spontaneously or were induced by IM or by Bmp7+Rosi. Iso was added to stimulate thermogenesis at the end of the culture period. Data are presented as mean ± SEM. ****, p < 0.0001; ***, p < 0.001; **, p < 0.01; *, p < 0.05; ns = nonsignificant. n = 3-5.

Article Snippet: The antibodies used for western blotting included PPARγ (Cell Signaling, #2430S, 1:1000 or Santa Cruz, #sc-7273, 1:500), FABP4 (Abcam, 13979, 1:2000), UCP1 (Abcam, #10983, 1:2000), BMP7 (Cell Signaling, #4693S, 1:1000), BMPR1A (Proteintech, #12701-1-AP, 1:1000), BMPR2 (Cell Signaling, #6979S, 1:1000), SMAD1 (Cell Signaling, #9743S, 1:1000) and β-ACTIN (Cell Signaling, #4976S, 1:2000), and HRP (Jackson ImmunoResearch, Code#715-035-150, 1:3000 or Code#715-035-152, 1:3000).

Techniques: Staining, Fluorescence, Isolation

Journal: World Journal of Stem Cells

Article Title: Bone marrow mesenchymal stem cell-derived exosomal lactate dehydrogenase A promotes tendon-bone healing via histone lactylation-mediated cartilage regeneration

doi: 10.4252/wjsc.v17.i9.106282

Figure Lengend Snippet: Lactate dehydrogenase A regulates H3K18 Lactylation and bone morphogenetic protein 7 expression in chondrocyte stem cells. A: Level of H3K18 Lactylation in chondrocyte stem cells from control, bone marrow mesenchymal stem cells and bone marrow mesenchymal stem cells + short hairpin RNA lactate dehydrogenase A groups; B: Enrichment of H3K18 Lactylation at the bone morphogenetic protein 7 (BMP7) promoter was measured by a chromatin immunoprecipitation-quantitative polymerase chain reaction assay; C: Promoter activity of BMP7 was analyzed by luciferase reporter gene assay; D and E: The RNA and protein level of BMP7 was analyzed by quantitative polymerase chain reaction and western blotting assay. a P < 0.05 vs control, b P < 0.01 vs control. LDHA: Lactate dehydrogenase A; BMSC-Exo: Exosomes derived from bone marrow mesenchymal stem cell; BMP: Bone morphogenetic protein 7.

Article Snippet: Primary antibodies included: LDHA (1:1000, Abcam ab52488, United Kingdom); H3K18 La (1:500, PTM Bio PTM-1401, China); BMP7 (1:1000, Santa Cruz sc-514365, TX, United States); p-Smad1/5/8 (1:1000, Cell Signaling 13820, MA, United States); CD63 (1:1000, System Biosciences EXOAB-CD63A-1, CA, United States); CD81 (1:1000, System Biosciences EXOAB-CD81A-1.

Techniques: Expressing, Control, shRNA, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Activity Assay, Luciferase, Reporter Gene Assay, Western Blot, Derivative Assay

Journal: Advanced Science

Article Title: WTAP Mediated m6A Modification Stabilizes PDIA3P1 and Promotes Tumor Progression Driven by Histone Lactylation in Esophageal Squamous Cell Carcinoma

doi: 10.1002/advs.202506529

Figure Lengend Snippet: BMP7 is a target of H4K8 lactylation in ESCC cells. A) The binding density of H4K8la was visualized by deepTools: the heatmap illustrates the CUT&Tag tag counts on the various H4K8la enrichment peaks in shNC and shPDIA3P1 cells. B) Distribution of H4K8la sites relative to translation start site (TSS). C Genome‐wide distribution of downregulated H4K8la‐binding peaks in shPDIA3P cells. D) KEGG analysis of the decreased H4K8la binding peaks at candidate target genes. E) Transcriptome sequencing was performed in control and shPDIA3P1 cells. F) Venn Diagram of CUT&Tag, RNA‐seq, GEO, and pubmed database to identify the potential downstream targets of H4K8la. G) Integrative Genomics Viewer tracks of CUT&Tag showing enriched H4K8la in the promotors of BMP7. The red rectangles indicate the peak regions of H4K8la on target‐gene promoters. H‐K) BMP7 mRNA and protein levels were measured in PDIA3P1‐KD cells (H, J) or PDIA3P1‐OE cells (I, K). L) Western blotting analysis of BMP7 expression in PDIA3P1‐KD cells cultured in lactate for 24 h. M) Western blot analysis of BMP7 expression in PDIA3P1‐OE cells cultured in 2‐DG or oxamate for 24 h. N, O) Using antibodies against H4K8la, CUT&Tag‐qPCR analysis for binding status at the BMP7 promotor of PDIA3P1‐KD cells (N) or PDIA3P1‐OE cells (O). These data represent the mean ± S.D. of triplicates. ** P < 0.01; *** P < 0.001.

Article Snippet: The IF primary antibody are shown as follows: GLUT1 (ET1601‐10, HUABIO), HK2 ( HA500186 , HUABIO), Flag (20543‐1‐AP, Proteintech), Pan‐Kla (HA722037, HUABIO), H4K8la (PTM‐1415RM, PTM‐BIO) and BMP7 (12221‐1‐AP, Proteintech).

Techniques: Binding Assay, Genome Wide, Sequencing, Control, RNA Sequencing, Western Blot, Expressing, Cell Culture

Journal: Advanced Science

Article Title: WTAP Mediated m6A Modification Stabilizes PDIA3P1 and Promotes Tumor Progression Driven by Histone Lactylation in Esophageal Squamous Cell Carcinoma

doi: 10.1002/advs.202506529

Figure Lengend Snippet: PDIA3P1 promotes tumorigenesis of esophageal squamous cell carcinoma through BMP7 both in vitro and in vivo. A) Western blot detection of BMP7 expression to reflect the effect of transfection of BMP7 by lentivirus. B‐G) Using lentivirus to construct BMP7 stable overexpression cell lines in TE‐1 and Eca‐109 cells with stably PDIA3P1‐KD. B) Proliferation of cells was analyzed using CCK8 assay. C) Tumor growth of TE‐1 and Eca‐109 cells was evaluated by colony formation assay. D) The proliferative abilities of ESCC cells were investigated using EdU assays. EdU scale bar: 100 µm. E) The flow cytometry showing apoptosis of cells by Annexin V‐APC and 7‐AAD staining. F) The migration and invasion ability was assessed by transwell assays. Transwell scale bar: 10 µm. G) Western blot shows expression levels of E‐Cadherin, N‐Cadherin, Vimentin, Snail, and BMP7. H‐K) Eca‐109 cells transfected with control, shPDIA3P1 or shPDIA3P1 and BMP7 were injected subcutaneously to establish a tumorigenesis model in nude mice ( n = 6). H) Photograph and comparison of tumor sizes in the indicated groups. I) The parameters of subcutaneous tumors were measured and recorded every 5 days, and calculated the volume of the tumor according to the formula below: tumor volume = 0.5 × length × width × width (mm3). J) Tumor weights in the indicated groups. K) Ki‐67 and BMP7 expression of representative IHC of nude mice tumor tissues. Scale bar, 20 µm. IF staining of TUNEL in subcutaneous tumor tissue sections. Scale bar: 50 µm. These data represent the mean ± S.D. of triplicates. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: The IF primary antibody are shown as follows: GLUT1 (ET1601‐10, HUABIO), HK2 ( HA500186 , HUABIO), Flag (20543‐1‐AP, Proteintech), Pan‐Kla (HA722037, HUABIO), H4K8la (PTM‐1415RM, PTM‐BIO) and BMP7 (12221‐1‐AP, Proteintech).

Techniques: In Vitro, In Vivo, Western Blot, Expressing, Transfection, Construct, Over Expression, Stable Transfection, CCK-8 Assay, Colony Assay, Flow Cytometry, Staining, Migration, Control, Injection, Comparison, TUNEL Assay