primary antibodies against cxcr4  (Proteintech)

Journal: Asia Pacific Allergy

Article Title: Identification and validation of autophagy-related genes and exploration of their relationship with disease severity in chronic rhinosinusitis with nasal polyps

doi: 10.5415/apallergy.0000000000000159

Figure Lengend Snippet: qRT-PCR and immunohistochemistry staining validate the expression levels of the central genes in clinical samples. (A–E) The mRNA expression levels of CXCR4, TNFSF11, HMOX1, APOE , and SPP1 . (F) Representative immunohistochemistry staining of CXCR4, HMOX1 , and SPP1 . (G–I) Quantitative summary of the mean intensity of CXCR4, HMOX1 , and SPP1 . qRT-PCR (control group: n=21; E-CRSwNP group: n=9; NE-CRSwNP group: n=12). Immunohistochemistry staining (control group: n=3; E-CRSwNP group: n=3; NE-CRSwNP group: n=3). Images were captured at ×400 magnification. Data are shown as means ± SEM. E-CRSwNP, eosinophilic chronic rhinosinusitis with nasal polyps; IOD, integrated optical density; NE-CRSwNP, noneosinophilic chronic rhinosinusitis with nasal polyps; qRT-PCR, quantitative real-time polymerase chain reaction. *P <0.05, **P <0.01, ****P <0.0001. ns, P >0.05.

Article Snippet: Subsequently, the sections were subjected to incubation with primary antibodies against CXCR4 (1:100; Wanleibio, Shenyang, China), HMOX1 (1:150; Wanleibio), and SPP1 (1:100; Wanleibio) at a temperature of 4 °C for the duration of the overnight period.

Techniques: Quantitative RT-PCR, Immunohistochemistry, Staining, Expressing, Control, Real-time Polymerase Chain Reaction

Journal: Asia Pacific Allergy

Article Title: Identification and validation of autophagy-related genes and exploration of their relationship with disease severity in chronic rhinosinusitis with nasal polyps

doi: 10.5415/apallergy.0000000000000159

Figure Lengend Snippet: Western blotting analysis and correlation analysis between the central genes and LC3B in CRSwNP. (A–C) Protein expression levels of CXCR4, HMOX1 , and SPP1 were assayed by western blot (control group: n=4; E-CRSwNP group: n=4; NE-CRSwNP group: n=4). (D) The mRNA expression level of LC3B was assayed by qRT-PCR. (E) Protein expression levels of LC3B were assayed by western blot (control group: n=5; E-CRSwNP group: n=4; NE-CRSwNP group: n=4). (F–H) The correlation between CXCR4, HMOX1, SPP1 , and LC3B. The Pearson correlation test was used. qRT-PCR (control group: n=21; E-CRSwNP group: n=9; NE-CRSwNP group: n=12). Data are shown as means ± SEM. CRSwNP, chronic rhinosinusitis with nasal polyps; E-CRSwNP, eosinophilic chronic rhinosinusitis with nasal polyps; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; NE-CRSwNP, noneosinophilic chronic rhinosinusitis with nasal polyps; qRT-PCR, quantitative real-time polymerase chain reaction. *P <0.05, **P <0.01, ***P <0.001. ns, P >0.05.

Article Snippet: Subsequently, the sections were subjected to incubation with primary antibodies against CXCR4 (1:100; Wanleibio, Shenyang, China), HMOX1 (1:150; Wanleibio), and SPP1 (1:100; Wanleibio) at a temperature of 4 °C for the duration of the overnight period.

Techniques: Western Blot, Expressing, Control, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

Journal: Asia Pacific Allergy

Article Title: Identification and validation of autophagy-related genes and exploration of their relationship with disease severity in chronic rhinosinusitis with nasal polyps

doi: 10.5415/apallergy.0000000000000159

Figure Lengend Snippet: The correlation between CXCR4, HMOX1 , and SPP1 and the disease severity parameters. The Spearman rank test was used. CT, computed tomography; NE, neutrophil elastase; SNOT-22, The Sino-Nasal Outcome Test 22.

Article Snippet: Subsequently, the sections were subjected to incubation with primary antibodies against CXCR4 (1:100; Wanleibio, Shenyang, China), HMOX1 (1:150; Wanleibio), and SPP1 (1:100; Wanleibio) at a temperature of 4 °C for the duration of the overnight period.

Techniques: Computed Tomography

Journal: Cancer Science

Article Title: Blocking CXCR4 – CARM1 – YAP axis overcomes osteosarcoma doxorubicin resistance by suppressing aerobic glycolysis

doi: 10.1111/cas.16295

Figure Lengend Snippet: Mechanistic diagram of role of the CXCR4–CARM1–YAP axis in drug resistance within osteosarcoma.

Article Snippet: For immunohistochemistry, sections were blocked and incubated with primary antibodies against CXCR4 and CARM1 (diluted 1:100 in 3% BSA; Cell Signaling Technology) at room temperature for 1 h, followed by the application of biotinylated donkey anti‐rabbit IgG (diluted 1:150 in 3% BSA; ThermoScientific) and HRP‐conjugated streptavidin (diluted 1:200 in 3% BSA; BioLegend, USA).

Techniques:

Journal: Cancer Science

Article Title: Blocking CXCR4 – CARM1 – YAP axis overcomes osteosarcoma doxorubicin resistance by suppressing aerobic glycolysis

doi: 10.1111/cas.16295

Figure Lengend Snippet: CARM1 is predicted as the downstream molecule of CXCR4. (A) Heatmap of differentially expressed mRNA in MG63 cell lines with or without the transfection of CXCR4 siRNA. (B) Volcano plot of highly differentially expressed mRNA in MG63 cell lines with or without CXCR4 siRNA transfection. (C) Top 10 enriched upregulated KEGG pathways of CXCR4 siRNA‐transfected MG63 cell lines. (D) Top 10 enriched downregulated KEGG pathways of MG63 cell lines transfected with CXCR4 siRNA. (E) The Kaplan–Meier survival curves show the effect of CARM1 on the progression of osteosarcoma. (F) GSEA analysis indicated that a significant change in glycolysis/gluconeogenesis was induced by knocking down CXCR4 in MG63 cell lines. (G) GSEA analysis showed significantly higher enrichment in oxidative phosphorylation pathways in MG63 cell lines with CXCR4 knockdown. All data are presented as mean ± SD and p ‐values were calculated using two‐tailed Student's t ‐test or Wilcox Rank Sum test. The p ‐values are presented as follows: ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: For immunohistochemistry, sections were blocked and incubated with primary antibodies against CXCR4 and CARM1 (diluted 1:100 in 3% BSA; Cell Signaling Technology) at room temperature for 1 h, followed by the application of biotinylated donkey anti‐rabbit IgG (diluted 1:150 in 3% BSA; ThermoScientific) and HRP‐conjugated streptavidin (diluted 1:200 in 3% BSA; BioLegend, USA).

Techniques: Transfection, Phospho-proteomics, Knockdown, Two Tailed Test

Journal: Cancer Science

Article Title: Blocking CXCR4 – CARM1 – YAP axis overcomes osteosarcoma doxorubicin resistance by suppressing aerobic glycolysis

doi: 10.1111/cas.16295

Figure Lengend Snippet: CXCR4 promotes glycolysis and modulates drug resistance in osteosarcoma via CARM1. (A) Representative immunofluorescence images of colocalization of CXCR4 and CARM1 were collected from four patients 100 µm. (B) Representative immunofluorescence images of CARM1 (green) in MG63 cell lines and drug‐resistant MG63‐DoxR cell lines with or without CXCR4 knockdown or CXCR4 overexpression 10 µm. (C) Drug resistance results of MG63 cell lines treated with or without CXCR4 overexpressing plasmids and CARM1 siRNA. (D) Drug resistance results of MG63‐DoxR cell lines transfected with or without CXCR4 siRNA and oe‐CARM1 plasmids. in MG63 cells * vs NC; ## vs CXCR4 OE; in MG63‐Dox cells ### vs CXCR4 siRNA. (E) Cell apoptosis rate of MG63 cell lines with or without the transfection of CXCR4 siRNA and oe‐CARM1 plasmid was detected by flow cytometry with annexin V‐FITC/PI double staining using quantitative FACS analysis. * vs NC; # vs CXCR4 siRNA; (F) Quantification of apoptosis rate and statistical analysis of MG63 cell lines with or without CXCR4 siRNA and oe‐CARM1 plasmid transfection. (G) Western blotting of several glycolytic enzymes in MG63 cell lines treated with or without CXCR4 OE and CARM1 siRNA. (H) Quantification of western blotting results and the statistical analysis of (G), *vs NC; # vs CXCR4 OE. All data are presented as mean ± SD and p ‐values were calculated using the two‐tailed Student's t ‐test or Wilcox Rank Sum test. The p ‐values are presented as follows: * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: For immunohistochemistry, sections were blocked and incubated with primary antibodies against CXCR4 and CARM1 (diluted 1:100 in 3% BSA; Cell Signaling Technology) at room temperature for 1 h, followed by the application of biotinylated donkey anti‐rabbit IgG (diluted 1:150 in 3% BSA; ThermoScientific) and HRP‐conjugated streptavidin (diluted 1:200 in 3% BSA; BioLegend, USA).

Techniques: Immunofluorescence, Knockdown, Over Expression, Transfection, Plasmid Preparation, Flow Cytometry, Double Staining, Western Blot, Two Tailed Test

Journal: Cancer Science

Article Title: Blocking CXCR4 – CARM1 – YAP axis overcomes osteosarcoma doxorubicin resistance by suppressing aerobic glycolysis

doi: 10.1111/cas.16295

Figure Lengend Snippet: The CXCR4–CARM1 axis promotes the progression of osteosarcoma by regulating the methylation of H3R17me and stimulates the transcription of YAP. (A) Protein levels of p‐YAP, YAP, and H3R17me in MG63 cells were analyzed under four conditions: Vector, CXCR4 OE, CARM1 siRNA, and CXCR4 OE + CARM1 siRNA. (B) Protein levels of p‐YAP, YAP, and H3R17me in MG63 cells were analyzed under four conditions: Vector, CXCR4 siRNA, CARM1OE, and CXCR4 siRNA + CARM1 OE. (C) Quantitative analysis of p‐YAP, YAP, and H3R17me in MG63 cell lines under different groups. (D) Quantitative analysis of protein intensity of p‐YAP, YAP, and H3R17me in MG63‐DoxR cell lines under different groups. (E) Chromatin immunoprecipitation assay kit (ChIP) analysis of the promoter site of YAP in drug‐resistant MG63 DoxR cell lines with or without overexpression of CARM1. All data are presented as mean ± SD and p ‐values were calculated using the two‐tailed Student's t ‐test or Wilcox Rank Sum test. The p ‐values are presented as follows: ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001. * vs Vector; # vs CXCR4 OE.

Article Snippet: For immunohistochemistry, sections were blocked and incubated with primary antibodies against CXCR4 and CARM1 (diluted 1:100 in 3% BSA; Cell Signaling Technology) at room temperature for 1 h, followed by the application of biotinylated donkey anti‐rabbit IgG (diluted 1:150 in 3% BSA; ThermoScientific) and HRP‐conjugated streptavidin (diluted 1:200 in 3% BSA; BioLegend, USA).

Techniques: Methylation, Plasmid Preparation, Chromatin Immunoprecipitation, Over Expression, Two Tailed Test

Journal: Cancer Science

Article Title: Blocking CXCR4 – CARM1 – YAP axis overcomes osteosarcoma doxorubicin resistance by suppressing aerobic glycolysis

doi: 10.1111/cas.16295

Figure Lengend Snippet: CXCR4–CARM1 axis promotes osteosarcoma progress and modulates drug resistance in vivo . (A) Overexpression of CARM1 effectively promoted the volume of the tumor and CXCR4 antagonist, AMD3100 inhibited the proliferation of osteosarcoma in the MG63 DoxR‐bearing model. (B) The tumor volume was monitored every 3 days, and a tumor growth curve was generated. (C) Tumors were extracted and weighed after 32 days. (D) Protein levels of CXCR4, CARM1, H3R17me, YAP, HK2, PKM2, LDHA in the tumors from control, CARM1 OE, AMD3100 and CARM1 OE + AMD3100. (E) Relative intensity of protein levels of CXCR4, CARM1, H3R17me, YAP, HK2, PKM2, LDHA in the tumors from control, CARM1 OE, AMD3100 and CARM1 OE + AMD3100. (F) Representative immunohistochemistry staining results of Ki‐67 and YAP of tumors from control, CARM1 OE, AMD3100 and CARM1 OE + AMD3100 200 µm (low magnification), 50 µm (high magnification). All data are presented as mean ± SD and p ‐values were calculated using the two‐tailed Student's t ‐test or Wilcox Rank Sum test. The p ‐values are presented as follows: ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001. * vs Control, # vs CARM1 OE.

Article Snippet: For immunohistochemistry, sections were blocked and incubated with primary antibodies against CXCR4 and CARM1 (diluted 1:100 in 3% BSA; Cell Signaling Technology) at room temperature for 1 h, followed by the application of biotinylated donkey anti‐rabbit IgG (diluted 1:150 in 3% BSA; ThermoScientific) and HRP‐conjugated streptavidin (diluted 1:200 in 3% BSA; BioLegend, USA).

Techniques: In Vivo, Over Expression, Generated, Control, Immunohistochemistry, Staining, Two Tailed Test