Journal: Scientific Reports

Article Title: High mobility group box-1 protein promotes astrocytic CCL5 production through the MAPK/NF-κB pathway following spinal cord injury

doi: 10.1038/s41598-024-72947-2

Figure Lengend Snippet: Changes in CCL5 and HMGB1 protein levels changes at lesion sites following rat SCI. (A) ELISA measurement of CCL5 protein levels at lesion sites following SCI at 0 d, 1 d, 4 d and 7 d, respectively. (B) Western blot analysis of HMGB1 expression following SCI at 0 d, 1 d, 4 d and 7 d. (C) Quantification data are shown in ( B ). Quantities were normalized to endogenous β-actin. (D) ELISA analysis of CCL5 protein levels at lesion sites at 0 d, 1 d, 4 d and 7 d following with or without intrathecal injection of 10 µl of an HMGB1 neutralizing antibody (HMGB1 Ab, 50 µg/kg). (E) Immunofluorescence staining revealed the colocalization of CCL5 with GFAP-positive astrocytes before or after SCI at 4 d with or without the intrathecal injection of 10 µl of HMGB1 Ab (50 µg/kg). The rectangle indicates the region magnified. Arrowheads indicate positive signals. n = 6. Scale bars, 50 μm in (E) . The experiments were performed in triplicate. The error bars represent the standard deviation (* P < 0.05, ** P < 0.01).

Article Snippet: The lysates were centrifuged at 12,000×g for 15 min. CCL5 levels (R&D systems, cat. no. MMR00) were assessed via ELISA kits according to the manufacturer’s directions.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Injection, Immunofluorescence, Staining, Standard Deviation

Journal: Scientific Reports

Article Title: High mobility group box-1 protein promotes astrocytic CCL5 production through the MAPK/NF-κB pathway following spinal cord injury

doi: 10.1038/s41598-024-72947-2

Figure Lengend Snippet: Examination of CCL5 production in astrocytes following stimulation with rHMGB1. ( A , B ) Purified primary astrocytes were stained with GFAP and Hoechst 33,342, and the purity was greater than 90%. Scale bar, 50 μm. ( C , D ) ELISA analysis of CCL5 in the lysates and supernatants of primary astrocytes following stimulation with 0–2.5 µg/mL rat recombinant HMGB1 (rHMGB1) for 24 h, respectively. ( E , F ) ELISA assay was used to determine the production of CCL5 in the lysates and supernatants following primary astrocyte treatment with 0.5 µg/ml rHMGB1 in the presence of 2.5 µg/ml HMGB1 Ab, respectively. The experiments were performed in triplicate. The error bars represent the standard deviation (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: The lysates were centrifuged at 12,000×g for 15 min. CCL5 levels (R&D systems, cat. no. MMR00) were assessed via ELISA kits according to the manufacturer’s directions.

Techniques: Purification, Staining, Enzyme-linked Immunosorbent Assay, Recombinant, Standard Deviation

Journal: Scientific Reports

Article Title: High mobility group box-1 protein promotes astrocytic CCL5 production through the MAPK/NF-κB pathway following spinal cord injury

doi: 10.1038/s41598-024-72947-2

Figure Lengend Snippet: Effects of RAGE, TLR-2, or TLR-4 interference on HMGB1-induced CCL5 production in astrocytes. ( A , B ) Cell lysates and supernatants were tested by ELISA for the production of CCL5, following astrocyte treatment with 0.5 µg/ml rHMGB1 in the presence of the RAGE inhibitor FPS-ZM1 (2 µM) for 24 h. ( D , E ) ELISA was used to determine CCL5 protein levels in lysates and supernatants from astrocytes after stimulation with 0.5 µg/ml rHMGB1 in the presence of the TLR2 inhibitor C29 (10 µM) for 24 h. ( G , H ) CCL5 levels in lysates and supernatants of astrocytes were determined by ELISA after the astrocytes were treated with the TLR4 inhibitor TAK-242 (2 µM) in the presence of 0.5 µg/ml rHMGB1 for 24 h. ( C , F , I ) CCK8 assay of the effects of FPS-ZM1, C29, or TAK-242 on the viability of astrocytes. The experiments were performed in triplicate. The error bars represent the standard deviation (* P < 0.05, ** P < 0.01).

Article Snippet: The lysates were centrifuged at 12,000×g for 15 min. CCL5 levels (R&D systems, cat. no. MMR00) were assessed via ELISA kits according to the manufacturer’s directions.

Techniques: Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Standard Deviation

Journal: Scientific Reports

Article Title: High mobility group box-1 protein promotes astrocytic CCL5 production through the MAPK/NF-κB pathway following spinal cord injury

doi: 10.1038/s41598-024-72947-2

Figure Lengend Snippet: Determination of CCL5 synthesis-related protein levels in astrocytes following stimulation with rHMGB1. (A) Western blot analysis of the phosphorylation of the ERK, JNK, P38 kinase and p65NF-κB proteins after astrocytes were treated with 0–2.5 µg/mL rHMGB1 for 24 h. (B-E) Quantification data are shown in (A). Quantities were normalized to endogenous β-actin. The experiments were performed in triplicate. The error bars represent the standard deviation (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: The lysates were centrifuged at 12,000×g for 15 min. CCL5 levels (R&D systems, cat. no. MMR00) were assessed via ELISA kits according to the manufacturer’s directions.

Techniques: Western Blot, Standard Deviation

Journal: Scientific Reports

Article Title: High mobility group box-1 protein promotes astrocytic CCL5 production through the MAPK/NF-κB pathway following spinal cord injury

doi: 10.1038/s41598-024-72947-2

Figure Lengend Snippet: Effects of the inhibition of MAPK/NF-κB signaling on the HMGB1-induced production of CCL5 from astrocytes. ( A , B ) ELISA was used to determine the effects of 10 µM ERK inhibitor PD98059, 10 µM JNK inhibitor SP6001251 or 10 µM P38 inhibitor SB203580 in the presence of 0.5 µg/ml rHMGB1 for 24 h on CCL5 protein production levels in lysates and supernatants of astrocytes. (C) Western blot analysis of the activation of p65NFκB protein after astrocyte treatment with 10 µM ERK inhibitor PD98059 or 10 µM JNK inhibitor SP6001251 for 24 h in the presence of 0.5 µg/ml rHMGB1. (D) Quantification data are shown in (C). Quantities were normalized to endogenous β-actin. ( E , F ) ELISA analysis of CCL5 in the lysates and supernatants of primary astrocytes following challenge with 10 µM SN50 for 24 h in the presence of 0.5 µg/ml rHMGB1, respectively. The experiments were performed in triplicate. The error bars represent the standard deviation (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: The lysates were centrifuged at 12,000×g for 15 min. CCL5 levels (R&D systems, cat. no. MMR00) were assessed via ELISA kits according to the manufacturer’s directions.

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay, Western Blot, Activation Assay, Standard Deviation

Journal: Scientific Reports

Article Title: High mobility group box-1 protein promotes astrocytic CCL5 production through the MAPK/NF-κB pathway following spinal cord injury

doi: 10.1038/s41598-024-72947-2

Figure Lengend Snippet: Effects of HMGB1-mediated astrocytic CCL5 on the migration of BV2 microglia cells or RAW 264.7 macrophage cells in vitro. (A) Illustration of the BV2 or RAW 264.7 cells with rCCL5 coculture model. (B) Transwell assay analysis of migration the ability of BV2 or RAW 264.7 cells co-incubated with 0.1 µg/ml rCCL5 for 48 h. ( C ) and ( D ) Quantification data are shown in (B). (E) Illustration of the BV2 or RAW 264.7 cells and ACM coculture model. (F) Migration assay of BV2 or RAW 264.7 cells cocultured with ACM. ACM were prepared from astrocytes exposed to 0.5 µg/ml rHMGB1 for 24 h, followed by the supernatant was collected and incubated with 2.5 µg/ml IgG or CCL5 Ab for 4 h. (G) and (H) Quantification data are shown in (F). Scale bars, 100 μm in ( B ) and ( F ). The experiments were performed in triplicate. The error bars represent the standard deviation (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: The lysates were centrifuged at 12,000×g for 15 min. CCL5 levels (R&D systems, cat. no. MMR00) were assessed via ELISA kits according to the manufacturer’s directions.

Techniques: Migration, In Vitro, Transwell Assay, Incubation, Standard Deviation

Journal: Scientific Reports

Article Title: High mobility group box-1 protein promotes astrocytic CCL5 production through the MAPK/NF-κB pathway following spinal cord injury

doi: 10.1038/s41598-024-72947-2

Figure Lengend Snippet: Effects of HMGB1-induced astrocytic CCL5 on microglia/macrophage polarization. ( A ) and ( B ) qRT-PCR was used to determine the expression of M1 markers ( CD86 , iNOS and TNF-a ) and M2 markers ( CD206 , Arg1 and Ym1 ) after BV2 or RAW 264.7 cells incubated with 0.1 µg/ml rCCL5 for 24 h. ( C ) and ( D ) The expression levels of M1 and M2 markers were determined by qRT-PCR following BV2 or RAW 264.7 cells cocultured with ACM for 24 h. ACM was prepared by astrocytes challenge with 0.5 µg/ml rHMGB1 for 24 h, after which the supernatant was collected and incubated with 2.5 µg/ml IgG or CCL5 Ab for 4 h. The experiments were performed in triplicate. The error bars represent the standard deviation (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: The lysates were centrifuged at 12,000×g for 15 min. CCL5 levels (R&D systems, cat. no. MMR00) were assessed via ELISA kits according to the manufacturer’s directions.

Techniques: Quantitative RT-PCR, Expressing, Incubation, Standard Deviation

Journal: Scientific Reports

Article Title: High mobility group box-1 protein promotes astrocytic CCL5 production through the MAPK/NF-κB pathway following spinal cord injury

doi: 10.1038/s41598-024-72947-2

Figure Lengend Snippet: Immunofluorescence of microglia/macrophage and locomotor function assessment after the inhibition of HMGB1 or CCL5 by neutralizing antibodies in rat SCI. ( A ) Immunofluorescence of IBA1- or CD68-positive cells at 4 d following SCI after neutralizing antibody application by intrathecal injection of 10 µl of neutralizing antibodies (HMGB1 Ab, 50 µg/kg), CCL5 (CCL5 Ab, 50 µg/kg) or normal rabbit IgG (IgG, 50 µg/kg). n = 6. Scale bars, 50 μm. ( B , C ) Quantitative analysis of the intensity of IBA1- or CD68-labeled cells in ( A) , respectively. (D) HE staining of the injured spinal cord at 21 d after administration of 10 µl of HMGB1, CCL5 neutralizing antibodies or normal rabbit IgG. n = 6. Scale bars, 500 μm. (E) Quantification data are shown in ( D) . (F) Basso, Beattie, and Bresnahan (BBB) locomotor scale scores for hindlimb motor function in rats at 0 d, 7 d, 14 d, 21 d and 28 d following the intrathecal administration of the HMGB1 Ab, CCL5 Ab or normal rabbit IgG. n = 6. The experiments were performed in triplicate. The error bars represent the standard deviation (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: The lysates were centrifuged at 12,000×g for 15 min. CCL5 levels (R&D systems, cat. no. MMR00) were assessed via ELISA kits according to the manufacturer’s directions.

Techniques: Immunofluorescence, Inhibition, Injection, Labeling, Staining, Standard Deviation

Journal: Scientific Reports

Article Title: High mobility group box-1 protein promotes astrocytic CCL5 production through the MAPK/NF-κB pathway following spinal cord injury

doi: 10.1038/s41598-024-72947-2

Figure Lengend Snippet: Mechanistic diagram of HMGB1-mediated astrocytic CCL5 contributes to microglia/macrophages activation and recruitment.

Article Snippet: The lysates were centrifuged at 12,000×g for 15 min. CCL5 levels (R&D systems, cat. no. MMR00) were assessed via ELISA kits according to the manufacturer’s directions.

Techniques: Activation Assay

Journal: Life Metabolism

Article Title: Blockade of Arf1-mediated lipid metabolism in cancers promotes tumor infiltration of cytotoxic T cells via the LPE-PPARγ-NF-κB-CCL5 pathway

doi: 10.1093/lifemeta/load036

Figure Lengend Snippet: The Arf1 inhibitors promote T-cell infiltration by upregulating CCL5 chemokine. (a) Chord diagram of gene correlation in the Arf1-ablated MYC-ON liver tumors ( n = 4 in each group). (b) qRT-PCR for chemokines in CT26 cells treated with DMSO, DU101, or DU102. (c) Serum CCL5 levels in the CT26 allografts with the indicated treatments ( n = 6 in each group). (d) CCL5 mRNA levels in the liver tumors of MYC-ON mice that were treated with DMSO, DU101, or DU102 ( n = 9 in each group). (e) Experimental design for the co-culture system. (f) FACS analysis of the migration of CD3 + T cells in CT26 cells that were treated with DMSO, DU101, or DU102. (g) The mRNA levels of CCL5 in CT26 cells with the indicated knockdowns and treatments. (h) FACS analysis of CD3 + T-cell migration in CT26 cells with the indicated knockdowns and treatments. (i and j) Tumor volumes (i) and tumor weights (j) of CT26 allografts with the indicated knockdowns and treatments ( n = 10 in each group). (k) Percentages of infiltrated CD3 + T cells in CT26 allografts with the indicated knockdowns and treatments ( n = 3 in each group). Data are shown as mean ± SEM. Student’s t test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. n.s., no significance.

Article Snippet: The level of chemokine CCL5 in cultured medium and mouse serum was detected by Mouse RANTES ELISA Kit (BOSTER, Cat# EK0495) according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, Co-Culture Assay, Migration

Journal: Life Metabolism

Article Title: Blockade of Arf1-mediated lipid metabolism in cancers promotes tumor infiltration of cytotoxic T cells via the LPE-PPARγ-NF-κB-CCL5 pathway

doi: 10.1093/lifemeta/load036

Figure Lengend Snippet: The Arf1 inhibitors activate the PPARγ pathway by inducing unsaturated fatty acid (LPE). (a) Heatmap of different lipid fractions in the Arf1-deficient CT26 cells. (b) The mRNA levels of downstream genes of PPARγ were detected in CT26 cells treated with different concentrations of LPE (0, 100, 200, 250, and 300 μmol/L for 24 h) by qRT-PCR. (c and d) The mRNA levels of PPARγ in CT26 cells (c) and MYC-ON liver tumors (d) treated with DMSO or Arf1 inhibitors were detected by qRT-PCR. (e) The CCL5 mRNA levels in CT26 cells treated with different concentrations of LPE were detected by qRT-PCR. (f) The CCL5 mRNA levels in CT26 cells with vesicle or GW9662 treatment were detected by qRT-PCR. (g) The CCL5 levels in cell medium collected from CT26 cells with the indicated treatments were determined by ELISA. (h) The percentages of the infiltrated CD3 + T cells in CT26 cells with the indicated treatments. Data are shown as mean ± SEM. Student’s t test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. n.s., no significance.

Article Snippet: The level of chemokine CCL5 in cultured medium and mouse serum was detected by Mouse RANTES ELISA Kit (BOSTER, Cat# EK0495) according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Life Metabolism

Article Title: Blockade of Arf1-mediated lipid metabolism in cancers promotes tumor infiltration of cytotoxic T cells via the LPE-PPARγ-NF-κB-CCL5 pathway

doi: 10.1093/lifemeta/load036

Figure Lengend Snippet: The Arf1 inhibitors promote T-cell tumor infiltration by activating the NF-κB-CCL5 pathway. (a) The phosphorylated p65 in CT26 cells treated with vesicle, DU101, or DU102 was detected by western blot analysis. (b) The phosphorylated p65 in MYC-ON liver tumors after DMSO, DU101, or DU102 treatment was detected by IHC staining. The right was the statistical analysis of p-p65 + cells per field. (c) The relative CCL5 enrichment in CT26 cells with the indicated treatments was analyzed by ChIP-PCR assay. (d) The CCL5 mRNA levels in CT26 cells with the indicated treatments were detected by qRT-PCR. (e) The CCL5 levels in the cell medium collected from CT26 cells with the indicated treatments were detected by the mouse CCL5 ELISA kit. (f) FACS analysis of the infiltrated CD3 + T cells in CT26 cells with the indicated treatments. (g and h) The proportions of the infiltrated CD3 + T cells (g) and CD3 + CCR5 + T (h) cells in Hepa1-6 cells with the indicated treatments were detected by FACS analysis. Data are shown as mean ± SEM. Student’s t test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. n.s., no significance.

Article Snippet: The level of chemokine CCL5 in cultured medium and mouse serum was detected by Mouse RANTES ELISA Kit (BOSTER, Cat# EK0495) according to the manufacturer’s instructions.

Techniques: Western Blot, Immunohistochemistry, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: BMC Cancer

Article Title: Exercise accelerates recruitment of CD8 + T cell to promotes anti-tumor immunity in lung cancer via epinephrine

doi: 10.1186/s12885-024-12224-7

Figure Lengend Snippet: Transcriptional targets were examined by RNA-seq in tumors after exercise. a The number of differentially expressed genes in the Ex and Ctrl mice were detected by RNA-seq. b Volcano plot of differentially expressed genes in Ex compared with Ctrl tumor tissues. Red dots represent upregulated genes, and blue dots represent downregulated genes. c Heat map of the differentially expressed genes in Ex compared with Ctrl tumor tissues. d Pathway enrichment analysis of significantly upregulated or downregulated genes. e-j RT-qPCR validates the gene expression of Cxcl10 , Cxcl12 , Cxcl14 , Ppbp , Pf4 and Ccl8 in the tumors each group ( n = 3) after tumor inoculation. k-l The expression levels of Cxcl10 and Cxcl12 were detected in the tumors in each group by ELISA ( n = 8). The results of e-l are presented as mean ± SEM. Statistical analysis was performed using two-tailed unpaired t tests. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The concentrations of Ccl5, Cxcl10 and Cxcl12 were measured in Lewis lung cancercellsculture supernatants and in the serum of mice using Ccl5 ELISA kit (Cusabio, CSB-E09256m), Cxcl10 ELISA kit (Cusabio, CSB-E08183m) and Cxcl12 (Cusabio, CSB-E04723m).

Techniques: RNA Sequencing, Quantitative RT-PCR, Gene Expression, Expressing, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: BMC Cancer

Article Title: Exercise accelerates recruitment of CD8 + T cell to promotes anti-tumor immunity in lung cancer via epinephrine

doi: 10.1186/s12885-024-12224-7

Figure Lengend Snippet: Exercise promotes CD8 + T cell recruitment with the assistance of Ccl5 and Cxcl10. a-d Correlation between Ccl5, Cxcl9, Cxcl10, and Cxcl11 mRNA level and CD8 + T cell using QUANTISEQ. Data were obtained from TIMER 2.0 ( http://timer.comp-genomics.org/ ). e-h RT - qPCR analyses of mRNA expression of chemokines in the tumors of Ctrl and Ex groups ( n = 3). i-j ELISA analysis of serum levels of Cxcl10 and Ccl5 in Ctrl and Ex mice ( n = 8). k-m RT-qPCR analyses of mRNA expression of cytokines and PD-L1 in the tumors of Ctrl and Ex mice ( n = 3). n Western blot analysis the expression of PD-L1 in the tumors of Ctrl and Ex mice ( n = 3). Full-length blots/gels are presented in Fig. 4n of source data. o Automatic quantification of PD-L1 IHC staining in indicated mice tumors ( n = 8). p IHC staining of PD-L1 on tumor sections from different groups ( n = 8). q Western blot analysis the expression of P53 in the tumors of Ctrl and Ex mice ( n = 3). Full-length blots/gels are presented in Fig. 4q of source data. The results of e-m and o are presented as mean ± SEM. Statistical analysis was performed using two-tailed unpaired t tests. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The concentrations of Ccl5, Cxcl10 and Cxcl12 were measured in Lewis lung cancercellsculture supernatants and in the serum of mice using Ccl5 ELISA kit (Cusabio, CSB-E09256m), Cxcl10 ELISA kit (Cusabio, CSB-E08183m) and Cxcl12 (Cusabio, CSB-E04723m).

Techniques: Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Immunohistochemistry, Two Tailed Test

Journal: BMC Cancer

Article Title: Exercise accelerates recruitment of CD8 + T cell to promotes anti-tumor immunity in lung cancer via epinephrine

doi: 10.1186/s12885-024-12224-7

Figure Lengend Snippet: Exercise inhibits tumor progression by upregulation of EPI levels. a Serum levels of EPI in Ctrl, Ex and Ex-post groups were detected by ELISA in tumor-bearing mice ( n = 8). b-d EPI mice receiving daily injections of EPI (0.5 mg/kg i.p.) ( n = 8) and TE mice with daily running for 14 days after tumor inoculation ( n = 8) compared with TC mice ( n = 8). b Representative images. c Tumor weights of indicated groups. d Tumor volumes of different groups. e IHC staining of CD8, CD4, CD3 and GZMB on tumor sections from indicated groups ( n = 8). f Automatic quantification of CD8, CD4, CD3 and GZMB IHC staining in indicated mice tumors ( n = 8). Data are presented as the mean ± SEM in each group. Statistical analysis of a was performed using two-tailed unpaired t tests. c , d and f were performed using one-way analysis of variance (ANOVA). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The concentrations of Ccl5, Cxcl10 and Cxcl12 were measured in Lewis lung cancercellsculture supernatants and in the serum of mice using Ccl5 ELISA kit (Cusabio, CSB-E09256m), Cxcl10 ELISA kit (Cusabio, CSB-E08183m) and Cxcl12 (Cusabio, CSB-E04723m).

Techniques: Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Two Tailed Test

Journal: BMC Cancer

Article Title: Exercise accelerates recruitment of CD8 + T cell to promotes anti-tumor immunity in lung cancer via epinephrine

doi: 10.1186/s12885-024-12224-7

Figure Lengend Snippet: EPI regulates changes in chemokine levels. a-f RT-qPCR analyses the mRNA expression of chemokines and cytokines in the tumors of TC and EPI mice ( n = 3). g-h ELISA analysis serum levels of Cxcl10 and Ccl5 in TC, TE and EPI treated mice ( n = 8). i-j RT-qPCR detect the mRNA levels of Cxcl10 and Ccl5 in LLC cells after treated with different concentrations of EPI (0, 5, 10, 20 µM). k-l Cxcl10 and Ccl5 in the supernatant of LLC cells were measured by ELISA. m RT-qPCR analyses the mRNA expression of CD274 in the tumors of control TC and EPI groups ( n = 3). n Western blot analyses PD-L1 in LLC cells after EPI treatment for 24 h at different concentration (0, 5, 10, 20 µM). Full-length blots/gels are presented in Fig. 6n of source data. o IHC staining of PD-L1 on tumor sections from indicated groups ( n = 8). p Automatic quantification of PD-L1 IHC staining in indicated mice tumors ( n = 8). q Graphical abstract of this study. Exercise-induced elevation of EPI is involved in regulating Ccl5 and Cxcl10 expression, subsequently promoting CD8 + T cells recruitment, and ultimately inhibiting tumor progression. Data are presented as the mean ± SEM in each group. Statistical analysis of a-f and m were performed using two-tailed unpaired t tests. g-l and p were performed using one-way analysis of variance (ANOVA). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The concentrations of Ccl5, Cxcl10 and Cxcl12 were measured in Lewis lung cancercellsculture supernatants and in the serum of mice using Ccl5 ELISA kit (Cusabio, CSB-E09256m), Cxcl10 ELISA kit (Cusabio, CSB-E08183m) and Cxcl12 (Cusabio, CSB-E04723m).

Techniques: Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Control, Western Blot, Concentration Assay, Immunohistochemistry, Two Tailed Test