DOI: 10.6084/m9.figshare.31230079
title: Data for Figure 3 - HGHJ medicated serum upregulates osteogenic gene expression (qRT-PCR raw Ct values of Runx2/Osterix/CollagenI/OPG/YAP/CTGF/GAPDH)

description: This dataset supports the core findings of the study titled "Henggu Gushang Heji Medicated Serum Enhances Osteogenic Potential of Rat Bone Marrow Mesenchymal Stem Cells through Modulation of the Hippo-YAP Pathway", focusing on the mechanistic role of a traditional Yi medicine formula (Henggu Gushang Heji, HGHJ) in regulating rat bone marrow mesenchymal stem cell (BMSC) function via the Hippo-YAP signaling pathway. The research falls within the interdisciplinary fields of Stem Cell Research​ and Cell Biology, with data types including Western blot full-membrane images, qRT-PCR raw Ct values, CCK-8 raw OD values, and Transwell migration raw images. 1. Research Background and Scientific Question​ Musculoskeletal disorders (e.g., fractures, osteoarthritis) pose global health challenges, with bone repair dependent on BMSC proliferation, migration, and osteogenic differentiation. HGHJ, a traditional Yi medicine formula documented in the Yunnan Provincial Drug Standards(2005), is clinically used for fracture and osteoarthritis management, but its cellular mechanisms for enhancing bone repair remain unclear. The Hippo-YAP pathway is a key regulator of stem cell behavior and osteogenesis; this study investigates whether HGHJ medicated serum enhances BMSC osteogenic potential by modulating this pathway. 2. Research Objectives​ Validate the effects of HGHJ medicated serum on BMSC proliferation, migration, and osteogenic differentiation. Elucidate HGHJ-mediated regulation of Hippo-YAP pathway components (YAP, p-YAP, LATS2, p-LATS2, CTGF). Determine the necessity of YAP signaling in HGHJ’s effects (via YAP inhibitor Verteporfin). 3. Experimental Design and Dataset Correspondence​ BMSCs were divided into four groups: control (10% blank serum), HGHJ (5% medicated serum), Verteporfin (YAP inhibitor), and combination (HGHJ + Verteporfin). Data were collected via four key experiments, with each dataset corresponding to specific figures and findings: Cell Proliferation (CCK-8 Assay): Raw OD values (including single-well measurements at 24h, 48h, and 72h for all groups) are provided. These data correspond to Figure 1, showing that the HGHJ group exhibited significantly higher OD at 72h (1.1581±0.06) compared to the control group (0.9672±0.05, p&lt;0.05), with Verteporfin partially reversing this effect. Cell Migration (Transwell Assay): Raw images (5 random fields per group, TIFF format) are included. These correspond to Figure 2, demonstrating that the HGHJ group had 2.2-fold higher migrated cells (229.3±13.7) than the control group (106.3±12.5, p&lt;0.001), while Verteporfin suppressed migration. Osteogenic Gene Expression (qRT-PCR): Raw Ct values (3 replicates for Runx2, Osterix, Collagen I, OPG, YAP, CTGF, and GAPDH) are provided. These correspond to Figure 3, showing reduced Ct values for Runx2, Osterix, Collagen I, and OPG in the HGHJ group (p&lt;0.05), indicating upregulated gene expression. Protein Expression (Western Blot): Full-membrane images (10 targets: YAP, p-YAP, LATS2, p-LATS2, CTGF, Runx2, Osterix, Collagen I, OPG, β-actin) are included. These correspond to Figure 3 (osteogenic proteins) and Figure 4 (Hippo-YAP pathway), showing that HGHJ reduced YAP/p-YAP/LATS2 phosphorylation, increased CTGF/osteogenic protein expression, and that Verteporfin reversed these effects. 4. Key Conclusions​ HGHJ medicated serum promotes BMSC proliferation, migration, and osteogenic differentiation​ (validated by CCK-8, Transwell, and Alizarin Red S staining). HGHJ modulates the Hippo-YAP pathway by inhibiting YAP/LATS2 phosphorylation and upregulating CTGF, thereby enhancing osteogenic gene/protein expression. Verteporfin (YAP inhibitor) partially reverses HGHJ’s effects, confirming YAP signaling as a key mediator (with potential ancillary pathways). 5. Research Significance​ This study provides the first evidence that HGHJ medicated serum enhances BMSC osteogenic potential via Hippo-YAP pathway modulation, bridging traditional Yi medicine application with modern molecular mechanisms. It offers a scientific basis for HGHJ’s clinical use in bone repair and highlights the Hippo-YAP pathway as a target for stem cell-based bone regeneration. 6. Data Availability​ This dataset contains raw, unprocessed data: uncropped Western blot membranes, unnormalized qRT-PCR Ct values, single-well CCK-8 OD values, and unquantified Transwell fields. Experimental methods follow standard protocols (e.g., CCK-8 assay, Transwell migration, qRT-PCR, Western blotting), with statistical analysis via GraphPad Prism 8.0 (one-way ANOVA + Tukey’s post-hoc test; p&lt;0.05= significant). Associated Study: Henggu Gushang Heji Medicated Serum Enhances Osteogenic Potential of Rat Bone Marrow Mesenchymal Stem Cells through Modulation of the Hippo-YAP PathwayThis dataset supports the core findings of the study titled "Henggu Gushang Heji Medicated Serum Enhances Osteogenic Potential of Rat Bone Marrow Mesenchymal Stem Cells through Modulation of the Hippo-YAP Pathway", focusing on the mechanistic role of a traditional Yi medicine formula (Henggu Gushang Heji, HGHJ) in regulating rat bone marrow mesenchymal stem cell (BMSC) function via the Hippo-YAP signaling pathway. The research falls within the interdisciplinary fields of Stem Cell Research​ and Cell Biology, with data types including Western blot full-membrane images, qRT-PCR raw Ct values, CCK-8 raw OD values, and Transwell migration raw images.1. Research Background and Scientific Question​Musculoskeletal disorders (e.g., fractures, osteoarthritis) pose global health challenges, with bone repair dependent on BMSC proliferation, migration, and osteogenic differentiation. HGHJ, a traditional Yi medicine formula documented in the Yunnan Provincial Drug Standards(2005), is clinically used for fracture and osteoarthritis management, but its cellular mechanisms for enhancing bone repair remain unclear. The Hippo-YAP pathway is a key regulator of stem cell behavior and osteogenesis; this study investigates whether HGHJ medicated serum enhances BMSC osteogenic potential by modulating this pathway.2. Research Objectives​Validate the effects of HGHJ medicated serum on BMSC proliferation, migration, and osteogenic differentiation.Elucidate HGHJ-mediated regulation of Hippo-YAP pathway components (YAP, p-YAP, LATS2, p-LATS2, CTGF).Determine the necessity of YAP signaling in HGHJ’s effects (via YAP inhibitor Verteporfin).3. Experimental Design and Dataset Correspondence​BMSCs were divided into four groups: control (10% blank serum), HGHJ (5% medicated serum), Verteporfin (YAP inhibitor), and combination (HGHJ + Verteporfin). Data were collected via four key experiments, with each dataset corresponding to specific figures and findings:Cell Proliferation (CCK-8 Assay): Raw OD values (including single-well measurements at 24h, 48h, and 72h for all groups) are provided. These data correspond to Figure 1, showing that the HGHJ group exhibited significantly higher OD at 72h (1.1581±0.06) compared to the control group (0.9672±0.05, p&lt;0.05), with Verteporfin partially reversing this effect.Cell Migration (Transwell Assay): Raw images (5 random fields per group, TIFF format) are included. These correspond to Figure 2, demonstrating that the HGHJ group had 2.2-fold higher migrated cells (229.3±13.7) than the control group (106.3±12.5, p&lt;0.001), while Verteporfin suppressed migration.Osteogenic Gene Expression (qRT-PCR): Raw Ct values (3 replicates for Runx2, Osterix, Collagen I, OPG, YAP, CTGF, and GAPDH) are provided. These correspond to Figure 3, showing reduced Ct values for Runx2, Osterix, Collagen I, and OPG in the HGHJ group (p&lt;0.05), indicating upregulated gene expression.Protein Expression (Western Blot): Full-membrane images (10 targets: YAP, p-YAP, LATS2, p-LATS2, CTGF, Runx2, Osterix, Collagen I, OPG, β-actin) are included. These correspond to Figure 3 (osteogenic proteins) and Figure 4 (Hippo-YAP pathway), showing that HGHJ reduced YAP/p-YAP/LATS2 phosphorylation, increased CTGF/osteogenic protein expression, and that Verteporfin reversed these effects.4. Key Conclusions​HGHJ medicated serum promotes BMSC proliferation, migration, and osteogenic differentiation​ (validated by CCK-8, Transwell, and Alizarin Red S staining).HGHJ modulates the Hippo-YAP pathway by inhibiting YAP/LATS2 phosphorylation and upregulating CTGF, thereby enhancing osteogenic gene/protein expression.Verteporfin (YAP inhibitor) partially reverses HGHJ’s effects, confirming YAP signaling as a key mediator (with potential ancillary pathways).5. Research Significance​This study provides the first evidence that HGHJ medicated serum enhances BMSC osteogenic potential via Hippo-YAP pathway modulation, bridging traditional Yi medicine application with modern molecular mechanisms. It offers a scientific basis for HGHJ’s clinical use in bone repair and highlights the Hippo-YAP pathway as a target for stem cell-based bone regeneration.6. Data Availability​This dataset contains raw, unprocessed data: uncropped Western blot membranes, unnormalized qRT-PCR Ct values, single-well CCK-8 OD values, and unquantified Transwell fields. Experimental methods follow standard protocols (e.g., CCK-8 assay, Transwell migration, qRT-PCR, Western blotting), with statistical analysis via GraphPad Prism 8.0 (one-way ANOVA + Tukey’s post-hoc test; p&lt;0.05= significant).Associated Study: Henggu Gushang Heji Medicated Serum Enhances Osteogenic Potential of Rat Bone Marrow Mesenchymal Stem Cells through Modulation of the Hippo-YAP Pathway