cxcl12 recombinant mouse protein  (MedChemExpress)

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: Serum CXCL12 is significantly elevated in PNI patients at 72 hours post-operation. The expression level of CXCL12 in the serum was analyzed via ELISA. The data are presented as the means ± SDs, n=20, **** p < 0.0001

Article Snippet: Investigation of the effect of CXCL12 (Sino Biological, 50025-MNAE, China) on SCs ferroptosis and inflammatory response: SCs were divided into Control group, LPS group, and LPS + CXCL12 group.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: CXCL12 inhibits LPS-induced ferroptosis in SCs. ( A ) Effect of different concentrations of LPS on SC survival levels. ( B ) Effect of different concentrations of CXCL12 on SC survival levels after LPS-induced SC damage. ( C - E ) RT-qPCR analysis of mRNA expression levels of ferroptosis-related genes (ACSL4, GPX4, FSP1) in SCs after CXCL12 treatment. (F) Western blot analysis of the expression levels of ferroptosis-related proteins ACSL4, GPX4, and FSP1 in SCs after CXCL12 treatment. ( G - I ) Densitometric analysis of proteins ACSL4, GPX4, and FSP1. ( J , K ) Immunofluorescence analysis of the expression levels of ferroptosis-related proteins ACSL4 and GPX4 in SCs after CXCL12 treatment, Scale bar = 100 um. ( L , M ) Quantification of the relative fluorescence intensity of ACSL4 and GPX4. ( N , O ) Fluorescence images of ROS and Fe 2+ in SCs after CXCL12 treatment, Scale bar = 50 um. ( P , Q ) Quantification of the relative fluorescence intensity of ROS and Fe 2+ . ( R , S ) Expression levels of MDA and GSH in SCs after CXCL12 treatment. The data are presented as the means±SDs, n=3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, not significant

Article Snippet: Investigation of the effect of CXCL12 (Sino Biological, 50025-MNAE, China) on SCs ferroptosis and inflammatory response: SCs were divided into Control group, LPS group, and LPS + CXCL12 group.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Immunofluorescence, Fluorescence

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: CXCL12 activates the ERK/Nrf2 signaling pathway in SCs. ( A ) Western blot analysis of ERK, p-ERK, and Nrf2 protein expression levels in SCs after CXCL12 treatment. ( B - C ) Quantification of the gray values for the ERK, p-ERK, Total Nrf2 and Nuclear Nrf2 proteins. The data are presented as the means ± SDs, n=3, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Investigation of the effect of CXCL12 (Sino Biological, 50025-MNAE, China) on SCs ferroptosis and inflammatory response: SCs were divided into Control group, LPS group, and LPS + CXCL12 group.

Techniques: Western Blot, Expressing

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: CXCL12 inhibits SC ferroptosis via the ERK/Nrf2 signaling pathway. ( A ) Western blot analysis of Nrf2 expression levels in SCs after treatment with the ERK inhibitor U0126. ( B ) Densitometric analysis of Nrf2 protein. ( C ) Western blot analysis of the expression levels of ferroptosis-related proteins ACSL4, GPX4, and FSP1 in SCs after treatment with the ERK inhibitor U0126. ( D - F ) Densitometric analysis of proteins ACSL4, GPX4, and FSP1. The data are presented as the means ± SDs, n=3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, not significant

Article Snippet: Investigation of the effect of CXCL12 (Sino Biological, 50025-MNAE, China) on SCs ferroptosis and inflammatory response: SCs were divided into Control group, LPS group, and LPS + CXCL12 group.

Techniques: Western Blot, Expressing

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: CXCL12 inhibits NF-κB signaling pathway activation and inflammatory factor secretion in SCs. ( A ) Western blot analysis of NF-κB, p-NF-κB, IκBα, and p-IκBα protein expression levels in SCs after CXCL12 treatment. ( B - C ) Quantification of gray values for the NF-κB, p-NF-κB, κBα, and p-IκBα proteins. ( D - E ) Levels of IL-1β and TNF-α in SCs after CXCL12 treatment. The data are presented as the means ± SDs, n=3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Investigation of the effect of CXCL12 (Sino Biological, 50025-MNAE, China) on SCs ferroptosis and inflammatory response: SCs were divided into Control group, LPS group, and LPS + CXCL12 group.

Techniques: Activation Assay, Western Blot, Expressing

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: CXCL12 inhibits the inflammatory response in SCs via the NF-κB signaling pathway. ( A ) Western blot analysis of NF-κB, p-NF-κB, IκBα, and p-IκBα protein expression levels in SCs after NF-κB overexpression. ( B - C ) Quantification of the gray values for the NF-κB, p-NF-κB, IκBα, and p-IκBα proteins. ( D - E ) Levels of IL-1β and TNF-α in SCs after NF-κB overexpression. The data are presented as the means ± SDs, n=3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, not significant

Article Snippet: Investigation of the effect of CXCL12 (Sino Biological, 50025-MNAE, China) on SCs ferroptosis and inflammatory response: SCs were divided into Control group, LPS group, and LPS + CXCL12 group.

Techniques: Western Blot, Expressing, Over Expression

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: CXCL12 inhibits SC ferroptosis, thereby mitigating the inflammatory response. ( A ) Western blot analysis of the expression levels of ferroptosis-related proteins ACSL4, GPX4, and FSP1 in SCs after FAC treatment. ( B - D ) Densitometric analysis of proteins ACSL4, GPX4, and FSP1. ( E , F ) Fluorescence images of ROS and Fe 2+ in SCs after FAC treatment, Scale bar = 50µm. ( G , H ) Quantification of the relative fluorescence intensity of ROS and Fe 2+ . ( I , J ) Expression levels of MDA and GSH in SCs after FAC treatment. ( K ) Western blot analysis of the expression levels of NF-κB, p-NF-κB, IκBα, and p-IκBα in SCs after FAC treatment. ( L , M ) Densitometric analysis of proteins NF-κB, p-NF-κB, IκBα, and p-IκBα. ( N , O ) Expression levels of IL-1β and TNF-α in SCs after FAC treatment. The data are presented as the means ± SDs, n=3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, not significant

Article Snippet: Investigation of the effect of CXCL12 (Sino Biological, 50025-MNAE, China) on SCs ferroptosis and inflammatory response: SCs were divided into Control group, LPS group, and LPS + CXCL12 group.

Techniques: Western Blot, Expressing, Fluorescence

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: CXCL12 inhibits PNI-induced ferroptosis in SCs. (A) Western blot analysis of ferroptosis-related protein expression levels (ACSL4, GPX4, FSP1) in the sciatic nerve after CXCL12 treatment. (B-D) Quantification of the gray values for the ACSL4, GPX4, and FSP1 proteins. (E-F) Representative immunofluorescence images of the ferroptosis-related proteins ACSL4 and GPX4 in the sciatic nerve after CXCL12 treatment. Scale bar = 50 µm. (G, H) Quantification of the relative fluorescence intensities of ACSL4 and GPX4. (I-K) Levels of Fe2+, MDA, and GSH in the sciatic nerve after CXCL12 treatment. (L-M) Transmission electron microscopy images showing the condition of the mitochondria in the SCs of the sciatic nerve after CXCL12 treatment. The data are presented as the means ± SDs, n=3, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: Investigation of the effect of CXCL12 (Sino Biological, 50025-MNAE, China) on SCs ferroptosis and inflammatory response: SCs were divided into Control group, LPS group, and LPS + CXCL12 group.

Techniques: Western Blot, Expressing, Immunofluorescence, Fluorescence, Transmission Assay, Electron Microscopy

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: CXCL12 activates the ERK/Nrf2 signaling pathway in the sciatic nerve. ( A ) Western blot analysis of ERK, p-ERK, Total Nrf2 and Nuclear Nrf2 protein expression levels in the sciatic nerve after CXCL12 treatment. ( B - C ) Quantification of the gray values for the ERK, p-ERK, Total Nrf2 and Nuclear Nrf2 proteins. The data are presented as the means ± SDs, n=3, ** p < 0.01,*** p < 0.001, **** p < 0.0001

Article Snippet: Investigation of the effect of CXCL12 (Sino Biological, 50025-MNAE, China) on SCs ferroptosis and inflammatory response: SCs were divided into Control group, LPS group, and LPS + CXCL12 group.

Techniques: Western Blot, Expressing

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: CXCL12 inhibits ferroptosis in the sciatic nerve via the ERK/Nrf2 signaling pathway. ( A ) Western blot analysis of Nrf2 protein expression levels in the sciatic nerve after treatment with the ERK inhibitor U0126. ( B ) Quantification of the gray values for the Nrf2 protein. ( C ) Western blot analysis of ferroptosis-related protein expression levels (ACSL4, GPX4, FSP1) in the sciatic nerve after treatment with the ERK inhibitor U0126. ( D - F ) Quantification of the gray values for the ACSL4, GPX4, and FSP1 proteins. The data are presented as the means ± SDs, n=3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Investigation of the effect of CXCL12 (Sino Biological, 50025-MNAE, China) on SCs ferroptosis and inflammatory response: SCs were divided into Control group, LPS group, and LPS + CXCL12 group.

Techniques: Western Blot, Expressing

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: CXCL12 inhibits NF-κB signaling pathway activation and inflammatory factor secretion in the sciatic nerve. ( A ) Western blot analysis of NF-κB, p-NF-κB, IκBα, and p-IκBα protein expression levels in the sciatic nerve after CXCL12 treatment. ( B - C ) Quantification of the gray values for the NF-κB, p-NF-κB, IκBα, and p-IκBα proteins. ( D - E ) Levels of IL-1β and TNF-α in the sciatic nerve after CXCL12 treatment. The data are presented as the means ± SDs, n=3, **p < 0.01, ***p < 0.001, **** p < 0.0001

Article Snippet: Investigation of the effect of CXCL12 (Sino Biological, 50025-MNAE, China) on SCs ferroptosis and inflammatory response: SCs were divided into Control group, LPS group, and LPS + CXCL12 group.

Techniques: Activation Assay, Western Blot, Expressing

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: CXCL12 inhibits sciatic nerve ferroptosis, thereby mitigating the inflammatory response. ( A ) Western blot analysis of ferroptosis-related protein expression levels (ACSL4, GPX4, FSP1) in the sciatic nerve after FAC treatment. ( B - D ) Quantification of the gray values for the ACSL4, GPX4, and FSP1 proteins. ( E ‒ G ) Levels of Fe 2+ , MDA, and GSH in the sciatic nerve after FAC treatment. ( H ) Western blot analysis of NF-κB, p-NF-κB, IκBα and p-IκBα protein expression levels in the sciatic nerve after FAC treatment. ( I , J ) Quantification of the gray values for the NF-κB, p-NF-κB, IκBα and p-IκBα proteins. (K, L) Levels of IL-1β and TNF-α in the sciatic nerve after FAC treatment. The data are presented as the means ± SDs, n=3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Investigation of the effect of CXCL12 (Sino Biological, 50025-MNAE, China) on SCs ferroptosis and inflammatory response: SCs were divided into Control group, LPS group, and LPS + CXCL12 group.

Techniques: Western Blot, Expressing

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: CXCL12 promotes nerve regeneration after PNI. ( A ) PWT. ( B ) PWL. ( C ) Representative footprint images of rats post-surgery. ( D ) SFI. ( E ) H&E staining of the longitudinal section of the sciatic nerve. ( F ) Immunofluorescence images of MBP and NF200 in the sciatic nerve. ( G , H ) Quantification of the relative fluorescence intensity of proteins MBP and NF200. The data are presented as the means ± SDs, n=3, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Investigation of the effect of CXCL12 (Sino Biological, 50025-MNAE, China) on SCs ferroptosis and inflammatory response: SCs were divided into Control group, LPS group, and LPS + CXCL12 group.

Techniques: Staining, Immunofluorescence, Fluorescence

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Substrate stiffness modulates human gingival fibroblast paracrine signaling to promote osteogenic differentiation of human periodontal ligament cells

doi: 10.3389/fbioe.2026.1753774

Figure Lengend Snippet: Substrate stiffness regulates the behaviors of human gingival fibroblasts (HGFs). Real-time reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect gene expression levels of (A) anti-inflammatory markers, IL4 , and IL10 , (B) matrix metalloproteinase markers, including MMP9 , and TIMP1 , (C) chemokine, CXCL12 . The expression of GAPDH was used as an internal control. Data were statistically analyzed by one-way ANOVA followed by Tukey’s multiple comparison tests ( n = 3: P < 0.05). (D) The expression of GAPDH was used as an internal control. Data were statistically analyzed by one-way ANOVA followed by Tukey’s multiple comparison tests ( n = 3: P < 0.05). (D) The protein expression of CXCL12 was detected by ELISA analysis. Data were statistically analyzed by one-way ANOVA followed by Tukey’s multiple comparison tests ( n = 4: P < 0.05). Data are presented as the mean ± standard deviation (SD), with different letters indicating statistically significant differences between multiple groups. IL4, interleukin 4; IL10, interleukin 10; MMP9, matrix metalloproteinase 9; TIMP1, tissue inhibitor of matrix metalloproteinases 1; CXCL12, CXC motif chemokine 12; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PDMS, polydimethylsiloxane.

Article Snippet: CXCL12 protein in conditioned medium was measured by Human CXCL12/SDF-1α ELISA Kit (Quantikine, R&D systems.

Techniques: Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Expressing, Control, Comparison, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Substrate stiffness modulates human gingival fibroblast paracrine signaling to promote osteogenic differentiation of human periodontal ligament cells

doi: 10.3389/fbioe.2026.1753774

Figure Lengend Snippet: Substrate stiffness regulates human gingival fibroblasts (HGFs) behaviors under an inflammatory condition. Real-time RT-PCR was performed to detect gene expression levels of (A) IL4 and IL10 , (B) MMP9 and TIMP1 , and (C) CXCL12 . The expression of GAPDH was used as an internal control. Data were statistically analyzed by one-way ANOVA followed by Tukey’s multiple comparison tests ( n = 3: P < 0.05). (D) The expression protein of CXCL12 was detected by ELISA analysis. Data were statistically analyzed by one-way ANOVA followed by Tukey’s multiple comparison tests ( n = 4: P < 0.05). Data are presented as the mean ± standard deviation (SD), with different letters indicating statistically significant differences between multiple groups. IL4, interleukin 4; IL10, interleukin 10; MMP9, matrix metalloproteinase 9; TIMP1, tissue inhibitor of matrix metalloproteinases 1; CXCL12, CXC motif chemokine 12; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PDMS, polydimethylsiloxane; LPS, lipopolysaccharide.

Article Snippet: CXCL12 protein in conditioned medium was measured by Human CXCL12/SDF-1α ELISA Kit (Quantikine, R&D systems.

Techniques: Quantitative RT-PCR, Gene Expression, Expressing, Control, Comparison, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Substrate stiffness modulates human gingival fibroblast paracrine signaling to promote osteogenic differentiation of human periodontal ligament cells

doi: 10.3389/fbioe.2026.1753774

Figure Lengend Snippet: Mitogen-activated protein kinase (MAPK) pathway regulated substrate stiffness-induced CXCL12 expression in human gingival fibroblasts (HGFs). (A) Real-time RT-PCR was performed to detect gene expression levels of CXCL12 . The expression of GAPDH was used as an internal control. Data were statistically analyzed by one-way ANOVA followed by Tukey’s multiple comparison tests ( n = 3: P < 0.05). (B) The protein expression of CXCL12 was detected by ELISA analysis. Data were statistically analyzed by one-way ANOVA followed by Tukey’s multiple comparison tests. ( n = 4: P < 0.05). Data are presented as the mean ± standard deviation (SD), with different letters indicating statistically significant differences between multiple groups. CXCL12, CXC motif chemokine 12; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PDMS, polydimethylsiloxane.

Article Snippet: CXCL12 protein in conditioned medium was measured by Human CXCL12/SDF-1α ELISA Kit (Quantikine, R&D systems.

Techniques: Expressing, Quantitative RT-PCR, Gene Expression, Control, Comparison, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Substrate stiffness modulates human gingival fibroblast paracrine signaling to promote osteogenic differentiation of human periodontal ligament cells

doi: 10.3389/fbioe.2026.1753774

Figure Lengend Snippet: Effects of conditioned media of human gingival fibroblasts (HGF-CM) under different substrate stiffness on human periodontal ligament cells’ behaviors. (A) Human periodontal ligament cells (HPDLCs) cultured with HGF-CM in osteogenic medium for 24 h. HPDLCs morphology was demonstrated using a phase-contrast microscope. Scale bars: 300 μm. (B) Real-time RT-PCR was performed to detect gene expression levels of CXCR4 , which is receptor of CXCL12. (C) Real-time RT-PCR was performed to detect gene expression levels of pro-inflammatory cytokine, IL1b . (D) Real-time RT-PCR was performed to detect gene expression levels of MMP8 and TIMP1 . The expression of GAPDH was used as an internal control. (E) Immunofluorescence analysis was performed to detect the protein expression of CXCR4 (green). The cytoskeleton (F-actin; red) and nuclei (blue) were stained using rhodamine-phalloidin and DAPI, respectively. Scale bars: 50 μm. Data were statistically analyzed by one-way ANOVA followed by Tukey’s multiple comparison tests ( n = 3: P < 0.05). Data are presented as the mean ± standard deviation (SD), with different letters indicating statistically significant differences between multiple groups. IL1B, interleukin-1β; MMP8, matrix metalloproteinase 8; TIMP1, tissue inhibitor of matrix metalloproteinases 1; CXCR4, CXC motif receptor type 4; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PDMS, polydimethylsiloxane.

Article Snippet: CXCL12 protein in conditioned medium was measured by Human CXCL12/SDF-1α ELISA Kit (Quantikine, R&D systems.

Techniques: Cell Culture, Microscopy, Quantitative RT-PCR, Gene Expression, Expressing, Control, Immunofluorescence, Staining, Comparison, Standard Deviation

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Substrate stiffness modulates human gingival fibroblast paracrine signaling to promote osteogenic differentiation of human periodontal ligament cells

doi: 10.3389/fbioe.2026.1753774

Figure Lengend Snippet: Schematic illustration shows the role of human gingival fibroblast–conditioned media (HGF-CM) in regulating osteogenic differentiation of human periodontal ligament cells (HPDLCs). (A) ECM stiffness stimulates CXCL12 chemokine expression in HGFs, which is associated with enhanced osteogenic responses in HPDLCs. (B) A potential clinical application of this mechanism is the utilization of HGF-derived factors to suppress inflammatory bone resorption and stabilize periodontal tissues. CXCL12: CXC motif chemokine 12.

Article Snippet: CXCL12 protein in conditioned medium was measured by Human CXCL12/SDF-1α ELISA Kit (Quantikine, R&D systems.

Techniques: Expressing, Derivative Assay

Journal: Cancer Research

Article Title: SPP1 Drives Colorectal Cancer Liver Metastasis and Immunotherapy Resistance by Stimulating CXCL12 Production in Cancer-Associated Fibroblasts

doi: 10.1158/0008-5472.CAN-24-4916

Figure Lengend Snippet: SPP1 promotes colorectal cancer metastasis through a positive feedback loop mediated by CAF-secreted CXCL12. A, Mass spectrometry analyzed supernatants from SPP1-stimulated and unstimulated CAFs, showing fold changes in secreted proteins (SPP1/control). B, A bubble chart displays commonly secreted protein levels in fibroblasts. C and D, Uniform Manifold Approximation and Projection (UMAP) plots and quantitative analysis reveal CXCL12 expression in fibroblasts within OE-SPP1 and vector groups. E, ELISA measured CXCL12 in CAF supernatants with/without SPP1 (1 µg/mL), n = 3. F, A flowchart shows CAF-conditioned medium’s (CM) impact on colorectal cancer (CRC) cell migration and invasion. G and H, Transwell and wound healing assays evaluated the effects of CAF-conditioned media or CXCL12-neutralizing antibody (100 ng/mL) on colorectal cancer cell migration and invasion ( n = 3). I–K, Flowchart illustrating the effects of CXCL12 or neutralizing antibody treatment on the colorectal cancer cell migration and invasion, assessed via transwell and wound healing assays ( n = 3). L, The effect of CXCL12 (100 ng/mL) or a neutralizing antibody (100 ng/mL) on the epithelial–mesenchymal transition markers expression in the colorectal cancer cells was analyzed using Western blotting ( n = 3). M, Correlation analysis of CXCL12 with SPP1 and TGFB1 in the TCGA dataset. N and O, The effect of CXCL12 (100 ng/mL) or neutralizing antibody (100 ng/mL) on the SPP1 and TGFβ expression in the colorectal cancer cells was evaluated using Western blotting ( N ) or ELISA ( O ), n = 3. Results are presented as mean ± SEM. P values were calculated using a two-tailed unpaired Student t test ( E ), whereas one-way ANOVA was used for the other comparisons. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: Human recombinant SPP1 (HY- P70499 ) and CXCL12 (HY- P70469 ) proteins were obtained from MedChemExpress.

Techniques: Mass Spectrometry, Control, Expressing, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Migration, Western Blot, Two Tailed Test

Journal: Cancer Research

Article Title: SPP1 Drives Colorectal Cancer Liver Metastasis and Immunotherapy Resistance by Stimulating CXCL12 Production in Cancer-Associated Fibroblasts

doi: 10.1158/0008-5472.CAN-24-4916

Figure Lengend Snippet: SPP1 inhibits T-cell infiltration and cytotoxicity via CXCL12 secretion from CAFs. A, Schematic of the coculture system with PDOs, T cells, and CAFs. CRC, colorectal cancer; E:T, effector to target. B and C, Confocal microscopy assessing the effect of SPP1 overexpression on T-cell infiltration and cytotoxicity in PDOs with or without CAFs ( n = 3). D and E, Impact of rhSPP1 (1 µg/mL) or CXCL12-neutralizing antibody (100 ng/mL) on T-cell infiltration and cytotoxicity in PDOs ( n = 3). Results are presented as mean ± SEM. P values were determined using one-way ANOVA. *, P < 0.05; **, P < 0.01; ***, P < 0.001; n.s., nonsignificant. PI, propidium iodide.

Article Snippet: Human recombinant SPP1 (HY- P70499 ) and CXCL12 (HY- P70469 ) proteins were obtained from MedChemExpress.

Techniques: Confocal Microscopy, Over Expression

Journal: Cancer Research

Article Title: SPP1 Drives Colorectal Cancer Liver Metastasis and Immunotherapy Resistance by Stimulating CXCL12 Production in Cancer-Associated Fibroblasts

doi: 10.1158/0008-5472.CAN-24-4916

Figure Lengend Snippet: SPP1 activates the β-catenin/HIF1α axis in the CAFs to drive CXCL12 secretion. A, Western blotting assessed key signaling pathway in CAFs after 24 hours of SPP1 protein stimulation. B–E, β-catenin and HIF1α expressions were analyzed following SPP1 or conditioned medium treatments, including from SPP1-overexpressing or -knockdown cells. F–H, HIF1α degradation was evaluated with MSAB or si-CTNNB1 transfection after cycloheximide (CHX) treatment, and HIF1α levels were measured after MSAB (1 µmol/L) or MG132 (20 µmol/L) pretreatment. I and J, Coimmunoprecipitation examined the HIF1α and β-catenin interaction. K and L, Immunofluorescence and nuclear–cytoplasmic fractionation assays assessed HIF1α and β-catenin localization ( n = 3). Scale bar, 25 μm. M–O, CXCL12 levels in conditioned media were measured after SPP1 (1 µg/mL) or MSAB treatments (24 hours). P, Correlation analysis of HIF1α and CXCL12 expression in 50 CAF samples using transcriptome data. Q, Dual-luciferase assays evaluated CXCL12 promoter activity ( n = 3). R and S, T-cell migration and infiltration were analyzed with or without SPP1 protein or MSAB treatment, n = 3. Scale bar, 50 μm. Western blotting ( A–J and L ) and ELISA ( M–O ) were repeated three times, with data representative of three independent experiments. Results are presented as mean ± SEM. P values were determined by one-way ANOVA ( M –O , R , and S ) and two-tailed unpaired Student t test ( F , G , and Q ). *, P < 0.05; **, P < 0.01; ***, P < 0.001. R and S , Created with Figdraw.com .

Article Snippet: Human recombinant SPP1 (HY- P70499 ) and CXCL12 (HY- P70469 ) proteins were obtained from MedChemExpress.

Techniques: Western Blot, Knockdown, Transfection, Immunofluorescence, Fractionation, Expressing, Luciferase, Activity Assay, Migration, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Cancer Research

Article Title: SPP1 Drives Colorectal Cancer Liver Metastasis and Immunotherapy Resistance by Stimulating CXCL12 Production in Cancer-Associated Fibroblasts

doi: 10.1158/0008-5472.CAN-24-4916

Figure Lengend Snippet: Blocking the SPP1/CXCL12 axis alleviates immunosuppression in the liver microenvironment and augments the benefits of immunotherapy. A, Flowchart of the intrasplenic injection model of liver metastasis using OE-SPP1 MC38 cells ( i.s.v. , intrasplenic injection; i.p. , intraperitoneal injection). B–D, Representative tumor morphology, hematoxylin and eosin staining, liver weight, and tumor burden ( n = 5 mice/group). Scale bar, 1 mm. E and F, Flow cytometric analysis of IFNγ + CD8 + and GZMB + CD8 + T cells in liver metastases ( n = 5 mice/group). G, Flowchart of the cecal orthotopic injection model of liver metastasis in the NOG mice using HCT116-HM cells. H and I, Luciferase images and bioluminescence quantification of metastatic livers. J, Hematoxylin and eosin staining and the number of liver metastases ( n = 5 mice/group). K, ELISA analysis of IFNγ levels in liver metastases ( n = 5 mice/group). L–N, ELISA of SPP1 and CXCL12 in peripheral blood of responders ( n = 25) and nonresponders ( n = 12) in immunotherapy-treated colorectal cancer cohorts. O, Diagram of tumor-derived SPP1 activation of CAFs to promote immunotherapy resistance in CRLM. Data are presented as mean ± SEM. P values were determined using one-way ANOVA ( C–F , and I–K ) and two-tailed unpaired Student t test ( L and M ). *, P < 0.05; **, P < 0.01; ***, P < 0.001. O, Created in BioRender. Liu, F. (2025) https://BioRender.com/k7tx8am .

Article Snippet: Human recombinant SPP1 (HY- P70499 ) and CXCL12 (HY- P70469 ) proteins were obtained from MedChemExpress.

Techniques: Blocking Assay, Injection, Staining, Luciferase, Enzyme-linked Immunosorbent Assay, Derivative Assay, Activation Assay, Two Tailed Test

Journal: bioRxiv

Article Title: IgE-producing cells on the move: CCR2 is a key regulator of IgE + plasma cell migration

doi: 10.64898/2025.12.18.695109

Figure Lengend Snippet: (A) Switched tonsil B cells were stained for surface CXCR4 followed by intracellular for IgE and IgG1. The histogram shows CXCR4 expression on IgE + (red) and IgG1 + (blue) gated cells. The filled grey histogram represents the isotype control staining. The accompanying graph shows CXCR4 median fluorescence intensity (MFI) on the surface of IgE + and IgG1 + cells. (B) CXCL12 induced migration of IgE + and IgG1 + cells was assessed using the transwell assay. After 3h of migration, the number of migrating cells was quantified by flow cytometry. Migration of IgE + and IgG1 + cells in response to 300 ng/mL (30nM) of CXCL12 is shown as a percentage of the cells migrating in response to RPMI control. (C) Flow cytometry dot plots of the IgE + and IgG1 + gated GC-like B cells, a PC-like “plasmablast” and PCs. Representative histograms show the CXCR4 expression on each of these gated IgE + and IgG1 + cells. (D) Bar chart showing the CXCR4 expression (MFI) across different IgE + and IgG1 + cell populations. (E) Flow cytometry staining of IgE + and IgG1 + GC-like B cells, PC-like PBs and PC after 3h of migration to the bottom chamber of the transwell. (F) CXCL12 induced migration shown as a percentage of the cells migrating in response to RPMI control. Data are mean + s.d. Statistical analysis was performed using one-way ANOVA with Tukey’s multiple comparison test (D, F) or paired two-tailed t -test with Welch’s correction (A, B); *p< 0.05; **p< 0.01; ***p< 0.001; and ****p< 0.0001. Non-significant values are not shown.

Article Snippet: More specifically, we tested the migration of IgE + and IgG1 + cells in response to recombinant human CXCL12 (R&D systems; 300ng/mL), CCL2 (Biolegend; 10ng/mL, 100ng/mL and 300ng/mL) and CCL28 (R&D systems; 300ng/mL and 1.5ug/mL).

Techniques: Staining, Expressing, Control, Fluorescence, Migration, Transwell Assay, Flow Cytometry, Comparison, Two Tailed Test