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R&D Systems recombinant mouse cxcl12 protein
Recombinant Mouse Cxcl12 Protein, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 84 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cxcl12+protein/pm42021544-65-0-4?v=R%26D+Systems
Average 95 stars, based on 84 article reviews

recombinant mouse cxcl12 protein - by Bioz Stars, 2026-07

95/100 stars

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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

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

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

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

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

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

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

Techniques: Western Blot, Expressing

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

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

Techniques: Western Blot, Expressing

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

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

Techniques: Activation Assay, Western Blot, Expressing

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

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

Techniques: Western Blot, Expressing, Over Expression

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

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

Techniques: Western Blot, Expressing, Fluorescence

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.

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.

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

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

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

Techniques: Western Blot, Expressing

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

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

Techniques: Western Blot, Expressing

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

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

Techniques: Activation Assay, Western Blot, Expressing

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

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

Techniques: Western Blot, Expressing

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

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

Techniques: Staining, Immunofluorescence, Fluorescence

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.

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

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.

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

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.

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

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.

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

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.

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

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.

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

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.

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

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

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

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 .

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

(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.

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

ATM-3507 inhibits the migration of DLBCL cells toward CXCL12. (A, B) NU-DUL-1 or Toledo cells were pre-treated with ATM-3507 or 0.03% DMSO (equivalent to the highest ATM-3507 concentration) for 1 h before being added to the upper chamber of a Transwell. The lower chamber contained 100 nM CXCL12 plus the same concentration of ATM-3507 or DMSO as in the upper chamber. After 4 h, the number of cells that had migrated into the lower chamber was determined using flow cytometry. The data are expressed as a percent of the number of DMSO-treated cells that migrated into the bottom chamber. The 100% values (percent of DMSO-treated cells that migrated into the lower chamber) in individual experiments ranged from 15-30% for NU-DUL-1 cells and from 50-70% for Toledo cells. Each symbol is an independent experiment. Means ± SEM are shown for 3 independent experiments. (C) NU-DUL-1 or Toledo cells were treated with 3 µM ATM-3507 or 0.03% DMSO for 1 h or 5 h before quantifying cell surface levels of CXCR4 by flow cytometry. Means ± SEM are shown for 3 independent experiments. p-values were calculated using two-tailed paired t -tests.

Journal: Frontiers in Immunology

Article Title: The tropomyosin 3.1/3.2 inhibitor ATM-3507 alters B-cell actin dynamics and impairs the growth and motility of diffuse large B-cell lymphoma cell lines

doi: 10.3389/fimmu.2025.1668379

Figure Lengend Snippet: ATM-3507 inhibits the migration of DLBCL cells toward CXCL12. (A, B) NU-DUL-1 or Toledo cells were pre-treated with ATM-3507 or 0.03% DMSO (equivalent to the highest ATM-3507 concentration) for 1 h before being added to the upper chamber of a Transwell. The lower chamber contained 100 nM CXCL12 plus the same concentration of ATM-3507 or DMSO as in the upper chamber. After 4 h, the number of cells that had migrated into the lower chamber was determined using flow cytometry. The data are expressed as a percent of the number of DMSO-treated cells that migrated into the bottom chamber. The 100% values (percent of DMSO-treated cells that migrated into the lower chamber) in individual experiments ranged from 15-30% for NU-DUL-1 cells and from 50-70% for Toledo cells. Each symbol is an independent experiment. Means ± SEM are shown for 3 independent experiments. (C) NU-DUL-1 or Toledo cells were treated with 3 µM ATM-3507 or 0.03% DMSO for 1 h or 5 h before quantifying cell surface levels of CXCR4 by flow cytometry. Means ± SEM are shown for 3 independent experiments. p-values were calculated using two-tailed paired t -tests.

Article Snippet: Imaging medium (600 μL) with the same concentration of ATM-3507 or DMSO plus 100 nM recombinant human CXCL12 (R&D Systems, Minneapolis, MN, USA #460-SD) was added to the bottom chamber.

Techniques: Migration, Concentration Assay, Flow Cytometry, Two Tailed Test

ATM-3507 inhibits the 2D motility of DLBCL cells on FN. Toledo DLBCL cells were added to FN-coated coverslips in the presence of 100 nM CXCL12 and either 10 µM ATM-3507 or 0.1% DMSO. After a 1 h pre-treatment period, time-lapse images were acquired every 30 s for 1 h. Cell tracks were generated from the time-lapse videos. Representative videos of DMSO-treated cells ( <xref ref-type=

Supplementary Video 1 ) and ATM-3507-treated cells ( Supplementary Video 2 ) are in the Supplementary Material . (A) Individual cell tracks from a representative experiment. In each experiment, 40–100 cell tracks were analyzed per condition. (B) Compiled data from 3 independent experiments. Each symbol represents the median value from an individual experiment. Means ± SEM are shown. p-values were calculated using two-tailed paired t -tests. " width="100%" height="100%">

Journal: Frontiers in Immunology

Article Title: The tropomyosin 3.1/3.2 inhibitor ATM-3507 alters B-cell actin dynamics and impairs the growth and motility of diffuse large B-cell lymphoma cell lines

doi: 10.3389/fimmu.2025.1668379

Figure Lengend Snippet: ATM-3507 inhibits the 2D motility of DLBCL cells on FN. Toledo DLBCL cells were added to FN-coated coverslips in the presence of 100 nM CXCL12 and either 10 µM ATM-3507 or 0.1% DMSO. After a 1 h pre-treatment period, time-lapse images were acquired every 30 s for 1 h. Cell tracks were generated from the time-lapse videos. Representative videos of DMSO-treated cells ( Supplementary Video 1 ) and ATM-3507-treated cells ( Supplementary Video 2 ) are in the Supplementary Material . (A) Individual cell tracks from a representative experiment. In each experiment, 40–100 cell tracks were analyzed per condition. (B) Compiled data from 3 independent experiments. Each symbol represents the median value from an individual experiment. Means ± SEM are shown. p-values were calculated using two-tailed paired t -tests.

Techniques: Generated, Two Tailed Test

Tpm3.1/3.2 regulates actin organization and actin-dependent processes in B cells. Tpm3.1/3.2 dimers associate with actin filaments, form homopolymers along the filament, stabilize the actin filament, and recruit myosin II. Myosin II dimers can crosslink actin filaments and mediate actomyosin contractility. Inhibition of Tpm3.1/3.2 by ATM-3507 impairs multiple actin-dependent processes in B cells, including the assembly of a peripheral actin ring that drives BCR-induced cell spreading via the formation of protrusive lamellipodia, and the formation of actomyosin arcs (indicated by the yellow arrow) at the inner face of the peripheral actin ring. ATM-3507 also inhibits the growth of DLBCL cells as well as their CXCL12-dependent migration and motility. Created with BioRender.com.

Journal: Frontiers in Immunology

Article Title: The tropomyosin 3.1/3.2 inhibitor ATM-3507 alters B-cell actin dynamics and impairs the growth and motility of diffuse large B-cell lymphoma cell lines

doi: 10.3389/fimmu.2025.1668379

Figure Lengend Snippet: Tpm3.1/3.2 regulates actin organization and actin-dependent processes in B cells. Tpm3.1/3.2 dimers associate with actin filaments, form homopolymers along the filament, stabilize the actin filament, and recruit myosin II. Myosin II dimers can crosslink actin filaments and mediate actomyosin contractility. Inhibition of Tpm3.1/3.2 by ATM-3507 impairs multiple actin-dependent processes in B cells, including the assembly of a peripheral actin ring that drives BCR-induced cell spreading via the formation of protrusive lamellipodia, and the formation of actomyosin arcs (indicated by the yellow arrow) at the inner face of the peripheral actin ring. ATM-3507 also inhibits the growth of DLBCL cells as well as their CXCL12-dependent migration and motility. Created with BioRender.com.

Techniques: Inhibition, Migration

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