stromal derived factor 1 beta  (R&D Systems)

Journal: Bone Research

Article Title: Neonatal bone marrow interstitial fluid supports expansion and osteogenic ability of human bone marrow mesenchymal stromal cells

doi: 10.1038/s41413-025-00496-z

Figure Lengend Snippet: NBIF promotes CXCR4 expression and enhances hBMSC homing to bone marrow. a, b Quantitative RT-PCR analysis of CXCR4 expression in hBMSCs after 7-day culture ( a ) and in mouse primary mBMSCs after 2 passages (7 days) ( b ). Data [and also in ( d )] were expressed as mean ± standard error of the mean (SEM) of the fold change across three replicates for each group. P -values were obtained from an unpaired t -test; ** P ≤ 0.01, **** P ≤ 0.000 1. c, d Representative images ( c ) and quantification data ( d ) of migratory hBMSCs in the Transwell culture (see the Materials and Methods section for details). rhCXCL12, recombinant human CXCL12 protein; AMD3100, the CXCR4 antagonist. Scale bar, 100 μm. e Scheme of the experimental design for mouse transplantation and analysis of GFP-labeled hBMSCs. f Quantification of the proportion of GFP + -hBMSCs in the whole bone marrow of host mice. Data were expressed as mean ± standard error of the mean (SEM) across indicated replicates for each group. 5 mice for the FBS-fed hBMSCs group and 5 mice for NBIF-fed hBMSCs group at each time point. P -values were obtained from an unpaired t -test; * P < 0.05, ** P < 0.01. g Representative immunofluorescent images of markers at 14 days post-transplantation. Scale bar, 10 μm. h–j Quantification of GFP + - ( h ), LEPR + - ( i ) or LEPR + ; GFP + - cells ( j ) in FBS-fed hBMSCs group ( n = 5 mice) or NBIF-fed hBMSC group ( n = 10 mice) 14 days post transplantation. Data were expressed as mean ± standard error of the mean (SEM) for each group. P -values were obtained from an unpaired t -test; * P < 0.05, ** P < 0.01

Article Snippet: To assess the effect of CXCL12, recombinant human CXCL12 was added to the lower chamber at a concentration of 100 ng/mL, with or without 50 nmol/L CXCR4 inhibitor AMD3100 (Medchemexpress) for 24 hours.

Techniques: Expressing, Quantitative RT-PCR, Recombinant, Transplantation Assay, Labeling

Journal: Cell Death Discovery

Article Title: CXCL12 alone is enough to Reprogram Normal Fibroblasts into Cancer-Associated Fibroblasts

doi: 10.1038/s41420-025-02420-0

Figure Lengend Snippet: A Schematic diagram showing the generation of NFs treated with CAFs-CM. B Phase-contrast microscopy revealed the typical spindle-like features of fibroblasts in CAFs and NFs isolated from lung cancer tissues, Scale bar = 100 μm. C Western blot analysis showed the expression of the fibroblast markers (FAP and Vimentin), the epithelial markers (CK-19) and the endothelial marker (CD31) in CAFs and NFs. D The impact of NFs and CAFs on the migratory capabilities of H1299 and A549 was assessed using transwell assay. The results include representative images of cell migration counts (left) and statistical data (right). Data are presented as the mean ± SD of three biological replicates.*, p < 0.05; ** , p < 0.005. Scale bar = 200 μm. E Western blot analysis showed the expression of CXCL12, CXCR4, pSTAT3/STAT3, Vimentin, and α-SMA in NFs treated with DMEM or CAFs CM.

Article Snippet: Recombinant human CXCL12 protein was purchased from Sino Biological (Beijing, China).

Techniques: Microscopy, Isolation, Western Blot, Expressing, Marker, Transwell Assay, Migration

Journal: Cell Death Discovery

Article Title: CXCL12 alone is enough to Reprogram Normal Fibroblasts into Cancer-Associated Fibroblasts

doi: 10.1038/s41420-025-02420-0

Figure Lengend Snippet: A . Western blot analysis of the expression of CXCL12, p-STAT3/STAT3, Vimentin, and α-SMA in p53S-CAFs treated with DMEM and NFs treated with DMEM, p53 -/- fibroblast CM, and p53S-CAFs CM. B Immunofluorescence analysis of the expression of Vimentin and α-SMA in NFs treated with DMEM, p53 -/- fibroblast CM, and p53S-CAFs CM. C Calculation of the percentage of cells that incorporated EdU or the number of migration cells for Transwell migration assay in NFs alone and treated with p53S-CAFs-CM. *, p < 0.05. **, p < 0.005. D Transwell migration assay of lung cancer cells H1299 and A549 cultured with NFs CM or CEFs CM. Average migration ±SEM from three independent experiments performed( n = 3). **, p < 0.005. ***, p < 0.001.

Article Snippet: Recombinant human CXCL12 protein was purchased from Sino Biological (Beijing, China).

Techniques: Western Blot, Expressing, Immunofluorescence, Migration, Transwell Migration Assay, Cell Culture

Journal: Cell Death Discovery

Article Title: CXCL12 alone is enough to Reprogram Normal Fibroblasts into Cancer-Associated Fibroblasts

doi: 10.1038/s41420-025-02420-0

Figure Lengend Snippet: A Western blot analysis of the expression of CXCL12 in NFs and CAFs. B Correlation of CXCL12 expression with CAFs markers α-SMA and Vimentin ( P < 0.001, R > 0).

Article Snippet: Recombinant human CXCL12 protein was purchased from Sino Biological (Beijing, China).

Techniques: Western Blot, Expressing

Journal: Cell Death Discovery

Article Title: CXCL12 alone is enough to Reprogram Normal Fibroblasts into Cancer-Associated Fibroblasts

doi: 10.1038/s41420-025-02420-0

Figure Lengend Snippet: A Western blotting analysis was conducted on NFs that were either untreated or treated with CXCL12, p53S-CAFs-CM, or a combination of p53S-CAFs-CM and AMD3100. B EdU incorporation assay of NFs treated with DMEM, CXCL12, CXCL12 and AMD3100, p53S-CAFs CM, p53S-CAFs CM and AMD3100. Representative images (left) and statistics (right) of the percentage of cells that incorporated EdU were shown. **, p < 0.005. C Transwell assay of NFs treated with DMEM, CXCL12, CXCL12 and AMD3100, p53S-CAFs CM, p53S-CAFs CM and AMD3100. Representative images (left) and statistics (right) of migration assay were shown. **, p < 0.005. ***, p < 0.001. D Transwell co-culture assay was employed to evaluate the pro-migratory capacity of normal fibroblasts, CEFs-12 (normal fibroblasts treated with CXCL12), CEFs (normal fibroblasts treated with p53S-CAFs conditioned medium), and CEFs+AMD3100 towards lung cancer cells H1299 and A549. The results include representative images of cell migration (left) and statistical data (right). **, p < 0.005, ***, p < 0.0005. Scale bar = 200 μm. E The flowchart for the experimental design of xenograft tumor models. F Representative images show the xenograft tumors in the backs of nude mice formed by mixed subcutaneous injection of CEFs-12 (or NFs) and A549 cells. G Tumor growth graphs indicated the tumor volumes at different time course ( n = 3 or 4 mice per group). ** p < 0.01. H Precise weighing of the tumors was performed upon dissection, ** p < 0.01.

Article Snippet: Recombinant human CXCL12 protein was purchased from Sino Biological (Beijing, China).

Techniques: Western Blot, Transwell Assay, Migration, Co-culture Assay, Injection, Dissection

Journal: Cell Death Discovery

Article Title: CXCL12 alone is enough to Reprogram Normal Fibroblasts into Cancer-Associated Fibroblasts

doi: 10.1038/s41420-025-02420-0

Figure Lengend Snippet: A p53S-CAFs-derived CXCL12 activated STAT3 pathway. NFs were treated with CXCL12 (20 ng/ml, 24 hours). The protein levels of CXCL12, CXCR4, IL6, p-JAK2, JAK2, p-STAT3, STAT3, SHP2, α-SMA, vimentin, p-ERK, ERK, p-AKT and AKT were detected by Western blot. B Western blot analysis of CXCL12, p-STAT3, STAT3, α-SMA and Vimentin from NFs alone and treated with CXCL12, or CXCL12 and Stattic. C Proliferative ability of NFs treated with CXCL12, CXCL12 and Stattic (upper), or p53S-CAFs CM, p53S-CAFs CM and Stattic(down) were measured by EdU incorporation assay. **, p < 0.005. D Migratory ability of NFs treated with CXCL12, CXCL12 and Stattic (upper), or p53S-CAFs CM, p53S-CAFs CM and Stattic(down) were measured by transwell migration assay. **, p < 0.005. ***, p < 0.001. Scale bar = 200 μm. E NFs were treated with CXCL12 and then transfected with STAT3 siRNA or control siRNA. STAT3 and p-STAT3 expression was measured by Western blot. *, p < 0.05. **, p < 0.005. F Proliferative and Migratory ability were measured by EdU incorporation assay and transwell assay. **, p < 0.005. ***, p < 0.001. G CEFs (NFs were treated with CXCL12) were transfected with STAT3 siRNA or control siRNA. Transwell assays detected the migration ability of H1299 and A549 co-cultured with CEFs or CEFs-SiSTAT3. **, p < 0.005. ***, p < 0.001. Scale bar = 200 μm.

Article Snippet: Recombinant human CXCL12 protein was purchased from Sino Biological (Beijing, China).

Techniques: Derivative Assay, Western Blot, Transwell Migration Assay, Transfection, Control, Expressing, Transwell Assay, Migration, Cell Culture

Journal: Cell Death Discovery

Article Title: CXCL12 alone is enough to Reprogram Normal Fibroblasts into Cancer-Associated Fibroblasts

doi: 10.1038/s41420-025-02420-0

Figure Lengend Snippet: A Western blot analysis of the knockdown efficiency following the transfection of CXCL12 siRNA into p53S-CAFs. B Western blot assessment of the impact on the expression of CAFs marker proteins in normal fibroblasts by conditioned medium after CXCL12 knockdown in p53S-CAFs. C Immunofluorescence analysis was conducted on the expression of CAFs markers α-SMA and Vimentin in p53 +/+ fibroblasts treated with either p53S-CAFs+NC CM or p53S-CAFs+siCXCL12 CM, in addition to the percentage of EdU-incorporated positive population. * p < 0.05, ** p < 0.005. D The impact of CEFs and CEFs+siCXCL12 on the migratory capabilities of H1299 and A549 was assessed using transwell assay. The results include representative images of cell migration counts (left) and statistical data (right). * p < 0.05; ** p < 0.005. Scale bar = 200 μm.

Article Snippet: Recombinant human CXCL12 protein was purchased from Sino Biological (Beijing, China).

Techniques: Western Blot, Knockdown, Transfection, Expressing, Marker, Immunofluorescence, Transwell Assay, Migration