Journal: Cell Death & Disease

Article Title: GIPC2 regulation of the PKM2/SREBP1 signaling axis controls adipogenic differentiation of mesenchymal stem cells

doi: 10.1038/s41419-025-08088-9

Figure Lengend Snippet: a Schematic diagram of Co-immunoprecipitation (Co-IP) assay followed by mass analysis. b Mass spectrometry (MS) analysis identified PKM2 as a potential interacting partner of GIPC2. c Interaction between GIPC2 and PKM2 assessed using Co-IP assays. d Colocalization of GIPC2 and PKM2 in MSCs revealed through immunofluorescence staining. e Protein structure prediction and molecular docking analysis performed to evaluate the binding affinity between PKM2 and GIPC2. f – h Interaction between f PKM2 and GIPC2 (deletion mutant of GIPC2△PDZ), g PKM2 and GIPC2 (deletion mutant of GIPC2△GH1), and h PKM2 and GIPC2 (deletion mutant of GIPC2△GH2) assessed using Co-IP assays. Error bars: mean ± s.e.m.; Scale bar, 25 μm ( d ).

Article Snippet: The cells were subsequently incubated overnight at 4 °C with the specific primary antibodies diluted in blocking buffer: GIPC2 (Abcam, ab175272, 1:100), PKM2 (Cell Signaling Technology, CST, 46687S, 1:100), SREBP1(Abcam, ab28481, 1:200).

Techniques: Co-Immunoprecipitation Assay, Mass Spectrometry, Immunofluorescence, Staining, Binding Assay, Mutagenesis

Journal: Cell Death & Disease

Article Title: GIPC2 regulation of the PKM2/SREBP1 signaling axis controls adipogenic differentiation of mesenchymal stem cells

doi: 10.1038/s41419-025-08088-9

Figure Lengend Snippet: a Nuclear localization of PKM2 in GIPC2-overexpressing MSCs assessed using immunofluorescence analysis. b Evaluation of PKM2 levels in the nuclear and cytoplasmic fractions of GIPC2-overexpressing MSCs using WB analysis. c Transcript levels of the adipogenic markers PPARG , C/EBPΑ , and FABP4 in GIPC2-overexpressing UC-MSCs with or without β-elemene treatment assessed using qRT-PCR. d Protein levels of the adipogenic markers PPAR-γ, C/EBP-α, and FABP4 in GIPC2-overexpressing UC-MSCs with or without β-elemene treatment assessed using WB analysis. e Visualization of lipid droplet formation in GIPC2-overexpressing UC-MSCs treated with β-elemene following 15 days of adipogenic induction obtained using ORO staining. All experiments were conducted in triplicate unless otherwise indicated. Statistical analysis: For the data in ( a – e ), a two-sided t -test was used if the normality criteria were met; otherwise, the two-sided Mann‒Whitney U test was applied. Within each replicate, we analyzed ≥ 50 randomly selected cells across multiple imaging fields ( b ). Error bars: mean ± s.e.m.; Scale bar, 25 μm ( a ), 10 μm ( a , enlarged image), 50 μm ( e ). a , b “GIPC2” indicates MSCs overexpressing GIPC2, and “Control” indicates MSCs not overexpressing GIPC2. c – e “GIPC2” indicates MSCs overexpressing GIPC2, and “β-elemene” indicates GIPC2-overexpressing MSCs with β-elemene.

Article Snippet: The cells were subsequently incubated overnight at 4 °C with the specific primary antibodies diluted in blocking buffer: GIPC2 (Abcam, ab175272, 1:100), PKM2 (Cell Signaling Technology, CST, 46687S, 1:100), SREBP1(Abcam, ab28481, 1:200).

Techniques: Immunofluorescence, Quantitative RT-PCR, Staining, Imaging, Control

Journal: Cell Death & Disease

Article Title: GIPC2 regulation of the PKM2/SREBP1 signaling axis controls adipogenic differentiation of mesenchymal stem cells

doi: 10.1038/s41419-025-08088-9

Figure Lengend Snippet: a GSEA comparing gene sets involved in the adipocytokine signaling pathway, lipid homeostasis, positive regulation of transcription by RNA polymerase and transcription coregulator binding between GIPC2-overexpressing UC-MSCs treated with or without β-elemene after 7 days of adipogenic induction. b Network plot illustrating transcription factors (TFs) enriched with differentially expressed genes (DEGs) in GIPC2-overexpressing UC-MSCs treated with or without β-elemene, with green nodes representing TFs and blue nodes representing target DEGs. c qRT-PCR analysis of SREBP1 in GIPC2-overexpressing UC-MSCs following β-elemene or PKM2-IN-1 treatment. d , e WB and immunofluorescence analyses evaluating SREBP1 expression in GIPC2-overexpressing UC-MSCs following β-elemene treatment. f qRT-PCR analysis of ACLY, FASN, FDPS, FDFT1 , and HMGCS1 in GIPC2-overexpressing UC-MSCs following β-elemene treatment. Transcriptomic analysis was performed on three samples per group ( n = 3) in ( a ). All experiments were conducted in triplicate unless otherwise indicated. Statistical analysis: one-way ANOVA ( c ), two-sided t -test ( d – f ). Error bars: mean ± s.e.m.; Scale bar, 100 μm ( e ), 10 μm ( e , enlarged image). a , d – f “GIPC2” indicates UC-MSCs overexpressing GIPC2, and “β-elemene” indicates GIPC2-overexpressing UC-MSCs with β-elemene. c “GIPC2” indicates UC-MSCs overexpressing GIPC2, “β-elemene” indicates GIPC2-overexpressing UC-MSCs with β-elemene, and “PKM2-IN-1” indicates GIPC2-overexpressing MSCs with PKM2-IN-1.

Article Snippet: The cells were subsequently incubated overnight at 4 °C with the specific primary antibodies diluted in blocking buffer: GIPC2 (Abcam, ab175272, 1:100), PKM2 (Cell Signaling Technology, CST, 46687S, 1:100), SREBP1(Abcam, ab28481, 1:200).

Techniques: Binding Assay, Quantitative RT-PCR, Immunofluorescence, Expressing

Journal: Cell Death & Disease

Article Title: GIPC2 regulation of the PKM2/SREBP1 signaling axis controls adipogenic differentiation of mesenchymal stem cells

doi: 10.1038/s41419-025-08088-9

Figure Lengend Snippet: a WB analysis of the adipogenic markers PPAR-γ, C/EBP-α, and FABP4 in GIPC2-overexpressing MSCs treated with Fatostatin. b ORO staining of Fatostatin-treated GIPC2-overexpressing UC-MSCs to assess lipid droplet formation after 15 days of adipogenic induction. c WB analysis of nuclear proteins isolated from UC-MSCs undergoing adipogenic differentiation for 15 days was performed to assess the nuclear expression of SREBP1 and PKM2 during the differentiation process. d Mechanism schematic diagram of GIPC2 promoting adipogenic differentiation in MSCs via the PKM2-SREBP1 axis. Statistical analysis: one-way ANOVA ( c ), two-sided t -test ( a , b ). Error bars: mean ± s.e.m.; Scale bar, 50 μm ( b ). a , b “GIPC2” indicates UC-MSCs overexpressing GIPC2, and “Fatostatin” indicates GIPC2-overexpressing MSCs with Fatostatin.

Article Snippet: The cells were subsequently incubated overnight at 4 °C with the specific primary antibodies diluted in blocking buffer: GIPC2 (Abcam, ab175272, 1:100), PKM2 (Cell Signaling Technology, CST, 46687S, 1:100), SREBP1(Abcam, ab28481, 1:200).

Techniques: Staining, Isolation, Expressing