Journal: Oncotarget

Article Title: Pancreatic cancer-derived exosomes promote tumor metastasis and liver pre-metastatic niche formation

doi: 10.18632/oncotarget.18831

Figure Lengend Snippet: Confocal microscopy (lung, liver, spleen, brain) of PKH67-labeled Panc02 EXO and Panc02-H7 EXO tissue distribution (green) 24 hpi. (A) PKH-67-labeled liposomes served as controls (scale bar=100 μm). Histogram shows exosome tissue distribution quantification (n=5/group). CD45, p-Stat3, and CD11b IF staining in liver sections from controls (left) and mice treated with Panc02 EXOs(middle) or Panc02-H7 EXOs (right) for 12 d without tumor challenge. (B) Histogram shows infiltrating CD45 + cell quantification. FN and α-SMA IF staining in liver sections from controls (top) and mice treated with Panc02 EXOs(middle) or Panc02-H7 EXOs (bottom) for 12 d without tumor challenge. (C) Histogram shows infiltrating α-SMA + hStCs and FN expression quantification(400× magnification; n=5/group). Western blotting analysis showed upregulated S100A8 and S100A9 in livers treated with Panc02-H7-derived exosomes. Histogram shows expression of the three proteins in three groupsas determined by densitometric analysis (n=3/group). (D) Pancreatic cancer-derived exosomes induce MDSC accumulation in peripheral blood. (E) Representative flow cytometric plots (left) and quantification (right) of CD11b + GR1 + MDSCs (n=5/group). *P<0.05, **P<0.01,***P<0.001.

Article Snippet: Primary antibodies against fibronectin (1:100),α-SMA (1:50), S100A8 (1:50), S100A9 (1:50), and F4/80 (1:50) were purchased from Proteintech (Wuhan, China).

Techniques: Confocal Microscopy, Labeling, Liposomes, Staining, Expressing, Western Blot, Derivative Assay

Journal: Oncotarget

Article Title: Pancreatic cancer-derived exosomes promote tumor metastasis and liver pre-metastatic niche formation

doi: 10.18632/oncotarget.18831

Figure Lengend Snippet: IHC analysis and histopathological examination of macrophages (F4/80), hStCs (α-SMA), and neutrophils in liver metastatic niches of naïve mice and mice treated with PBS, Panc02 EXOs, or Panc02-H7 EXOs at 30d post-SOI (arrow shows neutrophils in liver) (A) . Representative histogram shows quantification of F4/80 + macrophages, α-SMA + hStCs, and neutrophils (B) . Identification of FN, S100A8, and S100A9 as inflammatory mediators, and collagen deposition in the liver metastatic niche (C) . Representative histogram shows FN and MTS quantification (D) . Representative histogram shows S100A8 and S100A9 quantification (E) . n=6/group.**P<0.01,***P<0.001.10 fields assessed per sample. FOV, field of view.

Article Snippet: Primary antibodies against fibronectin (1:100),α-SMA (1:50), S100A8 (1:50), S100A9 (1:50), and F4/80 (1:50) were purchased from Proteintech (Wuhan, China).

Techniques:

Journal: Scientific Reports

Article Title: S100A8 regulated by estrogen improves injured endometrial epithelium reconstruction by promoting tight junction formation and stromal cell transformation

doi: 10.1038/s41598-025-08530-0

Figure Lengend Snippet: Effects of estrogen treatment on S100A8 expression in endometrium and epithelium morphology. ( a ) S100A8 immunohistochemical staining before and after estrogen treatment (1d/7d); black arrows indicate S100A8-positive immune cells (S100A8-PICs).The blue arrow indicates the release of S100A8 from S100A8-PICs. Scale bar is 100 or 400 μm. ( b ) Rat endometrial hematoxylin–eosin staining before and after estrogen treatment (1d/7d); red arrows show disrupted cell junctions. Scale bar is 100 μm. ( c ) S100A8-PICs number in the endometrium (1/mm 2 ) before and 1 d/7d after estrogen treatment (n = 18, from six rats, the number was counted from three immunohistochemical images of each sample , mean ± SEM). ( d ) S100A8 expression in the epithelium before and 1 d/7d after estrogen treatment (n = 18, from six rats, the expression level was measured from three immunohistochemical images of each sample, mean ± SEM). * Results with P values < 0.05 were considered significant. ** Results with P values < 0.01 were considered very significant.

Article Snippet: The cultured cells were randomly divided into three groups according to the experimental design: the control, S100A8 (0.5 μg/mL, HY- P71275 ; MCE, Shanghai, China), and S100A8 + FPS-ZM 1 (5 μM, HY-19370; MCE, Shanghai, China) groups.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Scientific Reports

Article Title: S100A8 regulated by estrogen improves injured endometrial epithelium reconstruction by promoting tight junction formation and stromal cell transformation

doi: 10.1038/s41598-025-08530-0

Figure Lengend Snippet: Effects of uterine cavity administration of S100A8 on the distribution of S100A8-PICs and endometrial morphology. ( a ) Distribution of S100A8-positive cells in the endometrium of four different groups with estrogen treatment (control, injury, P407, and S100A8 + P407 groups). Black arrows show S100A8-PICs. S100A8-PICs were distributed around the glands and blood vessels in the control group. S100A8-PICs in the site of injury or adhesion in the injury group and in the P407 group. S100A8-PICs reverse migrated to the glands or blood vessels in the S100A8 + P407 group. Scale bar is 100 μm or 1 mm. ( b ) Endometrial thickness in the four groups (n = 15, from 5 rats; 3 position was measured randomly in an image of each rat, mean ± SEM). ( c ) The number of endometrial glands in each group (n = 10, from 10 rats, glands were counted from a uterine horn cross-section image at 40 × of each rat, mean ± SEM). ( d ) The interior wall integrity of uterine cavity in each group (n = 10, from 10 rats, data was measured from a uterine horn cross-section image at 40 × of each rat, mean ± SEM). ( e ) The number of endometrial crypts in each group (n = 10, from 10 rats, crypts were counted from a uterine horn cross-section image at 40 × of each rat, mean ± SEM). * Results with P values < 0.05 were considered significant. ** Results with P values < 0.01 were considered very significant.

Article Snippet: The cultured cells were randomly divided into three groups according to the experimental design: the control, S100A8 (0.5 μg/mL, HY- P71275 ; MCE, Shanghai, China), and S100A8 + FPS-ZM 1 (5 μM, HY-19370; MCE, Shanghai, China) groups.

Techniques: Control

Journal: Scientific Reports

Article Title: S100A8 regulated by estrogen improves injured endometrial epithelium reconstruction by promoting tight junction formation and stromal cell transformation

doi: 10.1038/s41598-025-08530-0

Figure Lengend Snippet: Effects of S100A8 on the distribution of Ki-67-positive proliferating cells in the uteri and the proliferation of endometrial cells. ( a ) Ki-67-positive proliferating cells were distributed in the endometrium or uterine interior wall in all groups (control, injury, P407, and S100A8 + P407 groups). Scale bar is 100 or 400 μm. ( b ) Number of Ki-67-positive cells in the endometrium of each group (n = 20, from ten rats, the number was counted from two immunohistochemical images of each sample, mean ± SEM). ( c ) Number of Ki-67-positive cells in the uterine interior wall of each group (n = 20, from ten rats, the number was counted from two immunohistochemical images of each sample, mean ± SEM). ( d ) Effects of S100A8 and its receptor inhibitor (FPS-ZM1) on the proliferation of cultured endometrial cells in vitro (n = 5, mean ± SEM). Scale bar is 100 μm. * Results with P values < 0.05 were considered significant. ** Results with P values < 0.01 were considered very significant.

Article Snippet: The cultured cells were randomly divided into three groups according to the experimental design: the control, S100A8 (0.5 μg/mL, HY- P71275 ; MCE, Shanghai, China), and S100A8 + FPS-ZM 1 (5 μM, HY-19370; MCE, Shanghai, China) groups.

Techniques: Control, Immunohistochemical staining, Cell Culture, In Vitro

Journal: Scientific Reports

Article Title: S100A8 regulated by estrogen improves injured endometrial epithelium reconstruction by promoting tight junction formation and stromal cell transformation

doi: 10.1038/s41598-025-08530-0

Figure Lengend Snippet: Effects of S100A8 on tight junctions of endometrial epithelium or endometrial cells. ( a ) Immunohistochemical staining showing Claudin-1 distribution in the endometrium of different groups (control, injury, P407, and P407 + S100A8 groups); black arrows show Claudin-1 accumulated below the cell membrane of the lateral contact zone between adjacent cells and red arrows show cells expressing Claudin-1 in the stroma. Scale bar is 100 μm. ( b ) Claudin-1 expression in the endometrium of different groups (n = 10, mean ± SEM). ( c ) Western blot showing ZO-1 expression in endometrial cells of three different groups (control, S100A8, and S100A8 + FPS-ZM1 groups, n = 3, mean ± SEM). Actin was used as the loading control. The full-length bands can be found in supplementary Fig. 1. The bands of ZO-1 and actin cropped from different parts of the same gel. ( d ) Immunofluorescence showing the co-expression of Claudin-1 (red) and ZO-1 (green) in the endometrial cells of the three groups (n = 5, mean ± SEM). The nucleus is stained blue. The scale bar is 200 μm. * Results with P values < 0.05 were considered significant. ** Results with P values < 0.01 were considered very significant.

Article Snippet: The cultured cells were randomly divided into three groups according to the experimental design: the control, S100A8 (0.5 μg/mL, HY- P71275 ; MCE, Shanghai, China), and S100A8 + FPS-ZM 1 (5 μM, HY-19370; MCE, Shanghai, China) groups.

Techniques: Immunohistochemical staining, Staining, Control, Membrane, Expressing, Western Blot, Immunofluorescence

Journal: Scientific Reports

Article Title: S100A8 regulated by estrogen improves injured endometrial epithelium reconstruction by promoting tight junction formation and stromal cell transformation

doi: 10.1038/s41598-025-08530-0

Figure Lengend Snippet: Effects of S100A8 on endometrial “double-featured” cells. ( a ) Vimentin (red) and ZO-1 (green) immunofluorescent double staining of cultured endometrial cells. Scale bar is 200 μm. ( b ) Western blot showing CK-18 and vimentin expression in endometrial cells from each group (control, S100A8, and FPS-ZM1 groups; n = 3, mean ± SEM). The full-length bands can be found in supplementary Fig. 1. Actin was used as the loading control. As the target proteins have a similar molecular weight to that of the loading control, the bands of CK18/vimentin and actin were cropped from different gels. ( c ) Immunofluorescence showing vimentin expression in endometrial cells of each group (n = 5, mean ± SEM). Scale bar is 200 μm. * Results with P values < 0.05 were considered significant. ** Results with P values < 0.01 were considered very significant.

Article Snippet: The cultured cells were randomly divided into three groups according to the experimental design: the control, S100A8 (0.5 μg/mL, HY- P71275 ; MCE, Shanghai, China), and S100A8 + FPS-ZM 1 (5 μM, HY-19370; MCE, Shanghai, China) groups.

Techniques: Double Staining, Cell Culture, Western Blot, Expressing, Control, Molecular Weight, Immunofluorescence

Journal: Inflammation

Article Title: Programmed death-ligand 1 (PD-L1) Modulates Chemokine Production Via the TLR4/TRAF6 Signaling Axis During LPS + IFN-γ-Induced Endotoxemia-mimicked Sepsis

doi: 10.1007/s10753-025-02394-2

Figure Lengend Snippet: PD-L1 mediates the regulation of chemokine biosynthesis in M1-polarized macrophages, as determined by RNA-seq. ( A ) PD-L1 NC /THP-1-M and PD-L1 HI /THP-1-M cells were treated with LPS (100 ng/mL) + IFN-γ (20 ng/mL) for 6 h and subjected to RNA sequencing analysis. The KEGG database was used to identify enriched signaling pathways whose activation significantly differed (up). Single-gene GSEA was conducted using the GSE57065 , GSE65682 , and GSE95233 datasets to identify the relationship between PD-L1 and chemokine signaling pathways (down). ( B ) Intersection analysis of upregulated genes between the PD-L1 HI /THP-1-M + LPS + IFN-γ group and PD-L1 NC /THP-1-M + LPS + IFN-γ group and between the PD-L1 NC /THP-1-M + LPS + IFN-γ group and PD-L1 NC /THP-1-M + PBS group via a Venn diagram, followed by further intersection with chemokine datasets, revealed three highly expressed PD-L1-mediated chemokines (CCL8, CXCL9, and CXCL10) under LPS + IFN-γ stimulation. ( C ) RNA sequencing analysis revealed the expression of CCL8, CXCL9 and CXCL10 upon PD-L1 overexpression. ( D ) PD-L1 NC /THP-1-M and PD-L1 HI /THP-1-M cells were treated with LPS + IFN-γ for 12 h, after which CCL8 and CXCL9 mRNA and protein levels were quantified ( n = 3 independent biological replicates). ( E ) Human monocytes were induced into macrophages, transfected with scramble siRNA and PD-L1 siRNA, and then treated with LPS + IFN-γ for 12 h. CCL8 and CXCL9 mRNA and protein levels were quantified ( n = 3 independent biological replicates). ( F ) PD-L1 NC /THP-1-M and PD-L1 KO /THP-1-M cells were treated with LPS + IFN-γ for 12 h, after which CCL8 and CXCL9 mRNA and protein levels were quantified ( n = 3 independent biological replicates). The data are presented as the mean ± SEM. Statistical analysis for D, E and F was performed by one-way ANOVA with LSD test for post hoc analyses. * p < 0.05

Article Snippet: CCL8 and CXCL9 concentrations in cell supernatants or plasma were quantified using ELISA kits (Boster Biological Technology; Wuhan, China) with absorbance measurements at 450 nm, following the manufacturer’s protocols.

Techniques: RNA Sequencing, Protein-Protein interactions, Activation Assay, Expressing, Over Expression, Transfection

Journal: Inflammation

Article Title: Programmed death-ligand 1 (PD-L1) Modulates Chemokine Production Via the TLR4/TRAF6 Signaling Axis During LPS + IFN-γ-Induced Endotoxemia-mimicked Sepsis

doi: 10.1007/s10753-025-02394-2

Figure Lengend Snippet: TLR4 acts as a critical upstream regulator of PD-L1 in modulating CCL8 and CXCL9 expression. ( A ) GSEA of RNA-seq data to investigate the relationship between PD-L1 expression and Toll-like receptor signaling pathway activation. ( B ) PD-L1 HI /THP-1-M and PD-L1 NC /THP-1-M cells were transfected with scramble siRNA or TLR4 siRNA, respectively, followed by 12 h of stimulation with LPS (100 ng/mL) + IFN-γ (20 ng/mL). The supernatants and cells were harvested for concurrent measurements of CCL8 and CXCL9 protein and mRNA levels via ELISAs and qRT‒PCR ( n =3 independent biological replicates). The data are presented as the mean ± SEM. Statistical analysis for B was performed by one-way ANOVA with LSD test for post hoc analyses. * p < 0.05

Article Snippet: CCL8 and CXCL9 concentrations in cell supernatants or plasma were quantified using ELISA kits (Boster Biological Technology; Wuhan, China) with absorbance measurements at 450 nm, following the manufacturer’s protocols.

Techniques: Expressing, RNA Sequencing, Activation Assay, Transfection

Journal: Inflammation

Article Title: Programmed death-ligand 1 (PD-L1) Modulates Chemokine Production Via the TLR4/TRAF6 Signaling Axis During LPS + IFN-γ-Induced Endotoxemia-mimicked Sepsis

doi: 10.1007/s10753-025-02394-2

Figure Lengend Snippet: PD-L1 binds to the TLR4 downstream molecule TRAF6 to regulate CCL8 and CXCL9 expression. ( A ) Venn diagram analysis of proteins coimmunoprecipitated with PD-L1 in THP-1-M cells stimulated with LPS (100 ng/mL) + IFN-γ (20 ng/mL) for 6 h via IP‒MS. After excluding proteins bound to the IgG control group, 289 PD-L1-specific interacting proteins were identified (up). Representative MS/MS spectrum showing peptide fragments of the TRAF6 protein identified in the immunoprecipitation complex (down). ( B ) THP-1-M (up) and human monocyte-derived (down) macrophages were treated with or without LPS+IFN-γ for 6 h and then harvested for Co-IP. TRAF6 protein levels were measured via Western blotting. ( C ) PD-L1 HI /THP-1-M and PD-L1 NC /THP-1-M cells were transfected with scramble siRNA or TRAF6 siRNA, respectively, followed by 12 hours of stimulation with LPS+IFN-γ. The supernatants and cells were harvested for concurrent measurements of CCL8 and CXCL9 protein and mRNA levels via ELISAs and qRT‒PCR ( n =3 independent biological replicates). ( D ) Structural models of full-length PD-L1 and TRAF6 predicted by AlphaFold (PAE plots were obtained from AlphaFold). ( E ) Molecular docking generic matching information. ( F ) A protein‒protein interaction cartoon model generated by PyMOL (left). Predicted binding sites between PD-L1 and TRAF6 (right). ( G ) PD-L1 NC /THP-1-M, PD-L1 HI /THP-1-M and PD-L1 Δ62-139 /THP-1-M cells were treated with LPS+IFN-γ for 12 h. The supernatants and cells were harvested for concurrent measurements of CCL8 and CXCL9 protein and mRNA levels via ELISAs and qRT‒PCR ( n =3 independent biological replicates). The data are presented as the mean ± SEM. Statistical analysis for C and G was performed by one-way ANOVA with LSD test for post hoc analyses. * p <0.05.

Article Snippet: CCL8 and CXCL9 concentrations in cell supernatants or plasma were quantified using ELISA kits (Boster Biological Technology; Wuhan, China) with absorbance measurements at 450 nm, following the manufacturer’s protocols.

Techniques: Expressing, Control, Tandem Mass Spectroscopy, Immunoprecipitation, Derivative Assay, Co-Immunoprecipitation Assay, Western Blot, Transfection, Generated, Binding Assay

Journal: Inflammation

Article Title: Programmed death-ligand 1 (PD-L1) Modulates Chemokine Production Via the TLR4/TRAF6 Signaling Axis During LPS + IFN-γ-Induced Endotoxemia-mimicked Sepsis

doi: 10.1007/s10753-025-02394-2

Figure Lengend Snippet: PD-L1 knockout attenuates septic mouse mortality and suppresses CCL8 and CXCL9 expression. ( A – C ) PD-L1 WT /mice and PD-L1 KO /mice were intraperitoneally injected with LPS (15 mg/kg) and IFN-γ (10 µg/kg) to induce endotoxemia-induced sepsis. ( B ) The survival rate was determined over a 60-hour observation period and is presented as a Kaplan‒Meier survival curve ( n = 15 mice per group). The results are presented as Kaplan–Meier survival curves. The results of the statistical analysis were analyzed with the log-rank test. * p < 0.05. ( C ) Peripheral blood was collected 12 h posttreatment for serum isolation and PBMC separation. CCL8 and CXCL9 protein levels in serum and their corresponding mRNA levels in PBMCs were subsequently analyzed ( n = 3 independent biological replicates). ( D ) Peritoneal macrophages (PMs) were extracted from PD-L1 wild-type and knockout mice and then treated with LPS (100 ng/mL) + IFN-γ (20 ng/mL) for 12 h. The cells were harvested for CCL8 and CXCL9 mRNA and protein detection by qRT‒PCR and ELISAs ( n = 3 independent biological replicates). (E) Peripheral blood monocytes were extracted from PD-L1 wild-type and knockout mice and then treated with LPS + IFN-γ for 12 h. The cells were harvested for CCL8 and CXCL9 mRNA and protein detection by qRT‒PCR and ELISAs ( n = 3 independent biological replicates). The data are presented as the mean ± SEM. Statistical analysis for C was performed by Student’s t test, and that for D and E was performed by one-way ANOVA with LSD test for post hoc analyses. * p < 0.05

Article Snippet: CCL8 and CXCL9 concentrations in cell supernatants or plasma were quantified using ELISA kits (Boster Biological Technology; Wuhan, China) with absorbance measurements at 450 nm, following the manufacturer’s protocols.

Techniques: Knock-Out, Expressing, Injection, Isolation

Journal: Acta Biochimica et Biophysica Sinica

Article Title: ZNF3 regulates proliferation, migration and invasion through MMP1 and TWIST in colorectal cancer

doi: 10.3724/abbs.2022187

Figure Lengend Snippet: Table 1 Sequences of primers used in this study

Article Snippet: For antigen retrieval, tissue sections were boiled in 0.01 M citrate solution (pH 6.0) and incubated with primary antibody against ZNF3 (1:200; Proteintech, Wuhan, China).

Techniques:

Journal: Acta Biochimica et Biophysica Sinica

Article Title: ZNF3 regulates proliferation, migration and invasion through MMP1 and TWIST in colorectal cancer

doi: 10.3724/abbs.2022187

Figure Lengend Snippet: ZNF3 is highly expressed in CRC tissues and cell lines (A) Data from the UCSC Xena database show the mRNA expression levels of ZNF3 in 9701 Pan-Cancer tissues and in 738 para-carcinoma tissues. (B) mRNA expression levels of ZNF3 in 51 para-carcinoma tissues and 380 CRC tissues from the UCSC Xena database. (C) Immunohistochemical analysis of ZNF3 in 10 normal colon tissues and 70 colon cancer tissues at 20× and 40× magnification. Scale bar =100 μm (up) and scale bar =50 μm (down). (D) The quantitative analysis results of C. (E) RT-qRCR analysis of ZNF3 transcriptional expression levels in human normal colon epithelial cells (NCM460) and CRC cell lines (HT-29, HCT116, SW480 and LoVo). Data were analyzed by independent samples t test. * P<0.05, *** P<0.001, and **** P<0.0001.

Article Snippet: For antigen retrieval, tissue sections were boiled in 0.01 M citrate solution (pH 6.0) and incubated with primary antibody against ZNF3 (1:200; Proteintech, Wuhan, China).

Techniques: Expressing, Immunohistochemical staining

Journal: Acta Biochimica et Biophysica Sinica

Article Title: ZNF3 regulates proliferation, migration and invasion through MMP1 and TWIST in colorectal cancer

doi: 10.3724/abbs.2022187

Figure Lengend Snippet: ZNF3 knockdown inhibits CRC cell proliferation (A,B) Western blot analysis was carried out to verify the efficacy of ZNF3 overexpression in LoVo cells and ZNF3 knockdown in SW480 cells. (C) ZNF3 overexpression does not affect the proliferation of LoVo cells. (D) ZNF3 silencing reduced the proliferation of SW480 cells. (E) Cell cycle analysis in ZNF3-knockdown SW480 cells. Three independent experiments were conducted. Data are shown as the mean±SD. ** P<0.01, and *** P<0.001.

Article Snippet: For antigen retrieval, tissue sections were boiled in 0.01 M citrate solution (pH 6.0) and incubated with primary antibody against ZNF3 (1:200; Proteintech, Wuhan, China).

Techniques: Knockdown, Western Blot, Over Expression, Cell Cycle Assay

Journal: Acta Biochimica et Biophysica Sinica

Article Title: ZNF3 regulates proliferation, migration and invasion through MMP1 and TWIST in colorectal cancer

doi: 10.3724/abbs.2022187

Figure Lengend Snippet: ZNF3 promotes CRC cell migration and invasion (A) Migration of LoVo cells was detected by Transwell assay (magnification, 100×). (B) Quantitative analysis of migrated cells is shown as the mean±SD. (C) Migration and invasion of SW480 cells was detected by Transwell assay (magnification, 100×). (D) Quantitative analysis of migrated cells is shown as the mean±SD. * P<0.05, ** P<0.01, and **** P<0.0001.

Article Snippet: For antigen retrieval, tissue sections were boiled in 0.01 M citrate solution (pH 6.0) and incubated with primary antibody against ZNF3 (1:200; Proteintech, Wuhan, China).

Techniques: Migration, Transwell Assay

Journal: Acta Biochimica et Biophysica Sinica

Article Title: ZNF3 regulates proliferation, migration and invasion through MMP1 and TWIST in colorectal cancer

doi: 10.3724/abbs.2022187

Figure Lengend Snippet: ZNF3 regulates the expressions of EMT-related markers (A) The mRNA expression levels of EMT-related markers ( Vimentin, TWIST, CK19, MMP1, and ZEB2) were decreased in ZNF3-silenced SW480 cells. (B) The protein expression levels of EMT-related markers (Vimentin, TWIST, and MMP1) were decreased in ZNF3-silenced SW480 cells. (C) MMP1 was overexpressed in ZNF3-silenced SW480 cells. (D) TWIST was overexpressed in ZNF3-silenced SW480 cells. * P<0.05, and ** P<0.01.

Article Snippet: For antigen retrieval, tissue sections were boiled in 0.01 M citrate solution (pH 6.0) and incubated with primary antibody against ZNF3 (1:200; Proteintech, Wuhan, China).

Techniques: Expressing

Journal: Acta Biochimica et Biophysica Sinica

Article Title: ZNF3 regulates proliferation, migration and invasion through MMP1 and TWIST in colorectal cancer

doi: 10.3724/abbs.2022187

Figure Lengend Snippet: Overexpression of MMP1 and TWIST promotes the proliferation of ZNF3 -knockdown SW480 cells (A,B) Effect of MMP1 on the proliferation of ZNF3-knockdown SW480 cells. (C,D) Effect of TWIST on the proliferation of ZNF3-knockdown SW480 cells. (E,F) Effect of MMP1 and TWIST overexpression on the cell cycle of ZNF3-knockdown SW480 cells. * P<0.05, and ** P<0.01.

Article Snippet: For antigen retrieval, tissue sections were boiled in 0.01 M citrate solution (pH 6.0) and incubated with primary antibody against ZNF3 (1:200; Proteintech, Wuhan, China).

Techniques: Over Expression, Knockdown

Journal: Acta Biochimica et Biophysica Sinica

Article Title: ZNF3 regulates proliferation, migration and invasion through MMP1 and TWIST in colorectal cancer

doi: 10.3724/abbs.2022187

Figure Lengend Snippet: Overexpression of MMP1 and TWIST increases the migration and invasion of ZNF3 -knockdown SW480 cells (A) Effects of MMP1 and TWIST on the migration and invasion of ZNF3-RNAi#1 SW480 cells (magnification, 100×). (B) Effects of MMP1 and TWIST on the migration and invasion of ZNF3-RNAi#2 SW480 cells (magnification, 100×). * P<0.05, ** P<0.01, *** P<0.001, and **** P<0.0001.

Article Snippet: For antigen retrieval, tissue sections were boiled in 0.01 M citrate solution (pH 6.0) and incubated with primary antibody against ZNF3 (1:200; Proteintech, Wuhan, China).

Techniques: Over Expression, Migration, Knockdown

Journal: Acta Biochimica et Biophysica Sinica

Article Title: ZNF3 regulates proliferation, migration and invasion through MMP1 and TWIST in colorectal cancer

doi: 10.3724/abbs.2022187

Figure Lengend Snippet: Schematic overview of ZNF3-mediated cellular malignant behaviors in CRC

Article Snippet: For antigen retrieval, tissue sections were boiled in 0.01 M citrate solution (pH 6.0) and incubated with primary antibody against ZNF3 (1:200; Proteintech, Wuhan, China).

Techniques:

Journal: Frontiers in immunology

Article Title: Extracellular proteins as potential biomarkers in Sepsis-related cerebral injury.

doi: 10.3389/fimmu.2023.1128476

Figure Lengend Snippet: FIGURE 6 Levels of IL-6, CSF3, MMP8, and S100A8 and ROC curves of S100A8 and MMP8 in the peripheral blood of SAE patients. (A) When compared to the non-SAE (n=10) group, the SAE patients (n=21) had higher serum levels of MMP8 and S100A8 (both p<0.0). (B) High diagnostic accuracy was shown for the identification of SAE by both S100A8 (AUC = 0.962, 95% CI = 0.9001-1.000) and MMP8 (AUC = 0.7905, 95% CI = 0.6245-0.9564), with S100A8 having the highest accuracy. (C, D) Correlation analysis of the four extracellular molecules with clinical indicators and prognostic markers. S100A8, MMP8, CSF3, and IL-6 protein levels in the peripheral blood of sepsis patients were significantly linked with GCS scores (all p<0.05). S100A8 levels and 28-day mortality were strongly associated (r=0.634, p<0.01), and IL-6 levels and duration of mechanical breathing were positively correlated (r=0.360, p=0.046). *p <0.05, **p <0.01, ****p <0.0001; ns, no significance.

Article Snippet: The levels of IL6, MMP8, CSF3, and S100A8 were determined by the following commercial ELISA kits, according to manufacturer’s recommendations: MMP8 (Cat.#CSB-E04680h, Cusabio Biotech Co. Ltd, Wuhan, China), S100A8 (Cat. #CSB-E11833h, Cusabio Biotech Co. Ltd, Wuhan, China), IL-6 (Cat. #CSB-E04638h, Cusabio Biotech Co. Ltd, Wuhan, China), and CSF3 (Cat. #EK0360, Boster Biotech Co. Ltd, USA).

Techniques: Diagnostic Assay

Journal: Frontiers in immunology

Article Title: Extracellular proteins as potential biomarkers in Sepsis-related cerebral injury.

doi: 10.3389/fimmu.2023.1128476

Figure Lengend Snippet: FIGURE 7 Levels of MMP8, and S100A8 in the peripheral blood and CSF of sepsis rats. (A)Timeline of experiment procedures. (B) The recognition index of novel items in the CLP model rats (n =11) was lower than that of the Sham group rats (p < 0.05 vs Sham, n = 8). (C) In the Barnes maze experiment, the CLP model rats (n =11) made significantly more mistakes before finding the target hole on postoperative day 9 and 10 compared to the Sham group rats (p < 0.01, p < 0.05 vs Sham, n = 8). (D) The CLP rats (n=14) exhibited significantly elevated serum levels of MMP8 (p<0.05) and CSF levels (p=0.0506) in comparison to the SHAM group (n=12). (E) When compared to the SHAM (n=14) group, the CLP rats (n=12) had higher serum levels of S100A8(p<0.01) and CSF levels (all p<0.0001). CLP, cecal ligation and puncture; NS., no significance; NOR, novel object recognition. *p <0.05; ****p <0.0001.

Article Snippet: The levels of IL6, MMP8, CSF3, and S100A8 were determined by the following commercial ELISA kits, according to manufacturer’s recommendations: MMP8 (Cat.#CSB-E04680h, Cusabio Biotech Co. Ltd, Wuhan, China), S100A8 (Cat. #CSB-E11833h, Cusabio Biotech Co. Ltd, Wuhan, China), IL-6 (Cat. #CSB-E04638h, Cusabio Biotech Co. Ltd, Wuhan, China), and CSF3 (Cat. #EK0360, Boster Biotech Co. Ltd, USA).

Techniques: Comparison, Ligation

Journal: Cell Reports Medicine

Article Title: Alarmin S100A8 imparts chemoresistance of esophageal cancer by reprogramming cancer-associated fibroblasts

doi: 10.1016/j.xcrm.2024.101576

Figure Lengend Snippet:

Article Snippet: Human S100A8/S100A9 ELISA Kit , Proteintech , Cat# KE00177.

Techniques: Virus, shRNA, Over Expression, Recombinant, Purification, Staining, Enzyme-linked Immunosorbent Assay, Sonication, Chromatin Immunoprecipitation, Software

Journal: Cells

Article Title: S100A8-Mediated NLRP3 Inflammasome-Dependent Pyroptosis in Macrophages Facilitates Liver Fibrosis Progression

doi: 10.3390/cells11223579

Figure Lengend Snippet: DAMP S100A8 along with NLRP3 inflammasome-dependent pyroptosis is positively related to the progression of liver fibrosis. ( A ) Representative IHC images for S100A8 and S100A9 in liver sections from liver fibrosis patients and HCs. ( B , C ) ELISA analyses for serum levels of S100A8 and S100A9 in liver fibrosis patients and HCs. ( D ) Comparison of serum S100A8 and S100A9 levels in liver fibrosis patients with different phases. ( E – G ) Distribution of serum GSDMD ( E ), IL-1β ( F ), and IL-18 ( G ) levels in liver fibrosis patients with different phases (F0–4). ( H – J ) Correlation between serum S100A8 levels and GSDMD ( H ), IL-1β ( I ) or IL-18 ( J ) levels in liver fibrosis patients. ( K ) Representative mouse liver morphology and staining with H&E and Sirius Red. ( L – O ) IHC staining of mouse liver sections for NLRP3, S100A8, and S100A9. Black scale bar: 100 µm; Red scale bar: 50 µm. ELISA analyses for serum levels of S100A8 ( L ), GSDMD ( M ), IL-1β ( n ), and IL-18 ( O ) in 4-, 6-, and 8 week-mouse models of liver fibrosis. *** p < 0.001.

Article Snippet: Detailed ELISA kits were as follows: Mouse S100A8 (E-EL-M3048, elabscience, Wuhan, Hubei, China), mouse IL-1β (VAL601, Novus, Littleton, CO, USA), mouse IL-18 (E-EL-M0730c, elabscience, Wuhan, Hubei, China), mouse GSDMD (JL46371-96T, JiangLai, Baoshan, Shanghai, China), human S100A8 (E-EL-H1289c, elabscience, Wuhan, Hubei, China), human IL-1β (Mengbio, Shapingba, Chongqing, China), human IL-18 (Mengbio, Shapingba, Chongqing, China), and human GSDMD (Mengbio, Shapingba, Chongqing, China).

Techniques: Enzyme-linked Immunosorbent Assay, Comparison, Staining, Immunohistochemistry

Journal: Cells

Article Title: S100A8-Mediated NLRP3 Inflammasome-Dependent Pyroptosis in Macrophages Facilitates Liver Fibrosis Progression

doi: 10.3390/cells11223579

Figure Lengend Snippet: S100A8-mediated NLRP3 inflammasome-dependent pyroptotic macrophage death amplify the activation of human hepatic stellate cells. ( A – C ) The qRT–PCR analysis for the mRNA levels of NLRP3, pro-IL-1β, and pro-IL-18 in THP-1 macrophages treated with 0, 2, 5 or 10 µg/mL rhS100A8 or 5 µg/mL GST for 24 h. ( D ) The protein levels of NLRP3, GSDMD, GSDMD P30, pro-IL-1β, mature IL-1β, and cleaved caspase-1 were detected by Western blot in THP-1 macrophages treated with 5 µg/mL GST or rhS100A8. The protein expression was quantified by densitometry and normalized to β-actin and are shown as fold changes relative to the GST group (right panel). ( E ) PI and active caspase-1 double staining of pyroptotic cell death by flow cytometry in THP-1 macrophages treated with 5 µg/mL GST or rhS100A8. ( F – I ) Western blot analysis ( F ) and qRT-PCR analysis ( G–I ) of COL1A1, α-SMA, and TGF-β in LX-2 cells exposed to CM from THP-1 macrophages that were treated with 0, 2, 5 or 10 µg/mL of rhS100A8 or 5 µg/mL GST. ( J ) Western blot analysis of COL1A1, α-SMA, and TGF-β in LX-2 cells exposed to CM from THP-1 macrophages that were treated with 5 µg/mL of rhS100A8 with or without 1 h of MCC950 pretreatment. The protein expression was quantified by densitometry and normalized to β-actin and are shown as fold changes relative to the GST group (right panel). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Detailed ELISA kits were as follows: Mouse S100A8 (E-EL-M3048, elabscience, Wuhan, Hubei, China), mouse IL-1β (VAL601, Novus, Littleton, CO, USA), mouse IL-18 (E-EL-M0730c, elabscience, Wuhan, Hubei, China), mouse GSDMD (JL46371-96T, JiangLai, Baoshan, Shanghai, China), human S100A8 (E-EL-H1289c, elabscience, Wuhan, Hubei, China), human IL-1β (Mengbio, Shapingba, Chongqing, China), human IL-18 (Mengbio, Shapingba, Chongqing, China), and human GSDMD (Mengbio, Shapingba, Chongqing, China).

Techniques: Activation Assay, Quantitative RT-PCR, Western Blot, Expressing, Double Staining, Flow Cytometry

Journal: Cells

Article Title: S100A8-Mediated NLRP3 Inflammasome-Dependent Pyroptosis in Macrophages Facilitates Liver Fibrosis Progression

doi: 10.3390/cells11223579

Figure Lengend Snippet: TLR4/NF-κB signaling cascade and ROS abundance are responsible for S100A8-induced NLRP3 inflammasome-dependent pyroptotic death in macrophages. ( A ) Western blot analysis of p65, p-p65, IKKα, and p-IKKα expression in THP-1 macrophages treated with GST-rhS100A8 or GST for 0, 30, 60 or 120 min. The protein expression was quantified by densitometry and normalized to β-actin and are shown as fold changes relative to the 0 min group (right panel). ( B – E ) THP-1 macrophages were exposed to 5 µg/mL rhS100A8 with or without 1 h of BAY 11-7082, TAK-242 or FPS-ZM1 pretreatment. The qRT-PCR analysis was performed to detect the mRNA levels of NLRP3 ( B ), pro-IL-1β ( C ), and pro-IL-18 ( D ). Western blot analysis was used to determine the protein expression of NLRP3, GSDMD, GSDMD P30, pro-IL-1β, mature IL-1β, and cleaved caspase-1 ( E ). The protein expression was quantified by densitometry and normalized to β-actin and are shown as fold changes relative to the GST group (right panel). ( F ) THP-1 macrophages were pretreated with TAK-242 or FPS-ZM1 for 1 h and then exposed to 5 µg/mL of rhS100A8. Western blot analysis was used to determine the expression of p-p65 and p-IKKα. The protein expression was quantified by densitometry and normalized to β-actin and are shown as fold changes relative to the GST group (right panel). ( G ) Flow cytometry analysis of ROS levels in THP-1 macrophages treated with rhS100A8 for 6 h. ( H ) THP-1 macrophages were exposed to 5 µg/mL of rhS100A8 with or without 1 h of DPI pretreatment. Protein expression levels of NLRP3, GSDMD, GSDMD P30, pro-IL-1β, and mature IL-1β were determined by Western blot. The protein expression was quantified by densitometry and normalized to β-actin and are shown as fold changes relative to the GST group (right panel); ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Detailed ELISA kits were as follows: Mouse S100A8 (E-EL-M3048, elabscience, Wuhan, Hubei, China), mouse IL-1β (VAL601, Novus, Littleton, CO, USA), mouse IL-18 (E-EL-M0730c, elabscience, Wuhan, Hubei, China), mouse GSDMD (JL46371-96T, JiangLai, Baoshan, Shanghai, China), human S100A8 (E-EL-H1289c, elabscience, Wuhan, Hubei, China), human IL-1β (Mengbio, Shapingba, Chongqing, China), human IL-18 (Mengbio, Shapingba, Chongqing, China), and human GSDMD (Mengbio, Shapingba, Chongqing, China).

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Flow Cytometry

Journal: Cells

Article Title: S100A8-Mediated NLRP3 Inflammasome-Dependent Pyroptosis in Macrophages Facilitates Liver Fibrosis Progression

doi: 10.3390/cells11223579

Figure Lengend Snippet: The potential predictive powers of S100A8, GSDMD, IL-1β, and IL-18 for the occurrence and severity of liver fibrosis. ( A ) ROC curves of serum S100A8, GSDMD, IL-1β, and IL-18 for distinguishing liver fibrosis patients from HCs. ( B ) ROC curve, of serum S100A8, GSDMD, IL-1β, and IL-18 for detecting moderate-to-severe liver fibrosis from no or mild liver fibrosis in liver fibrosis patients. ( C ) A working model illustrating that S100A8-mediated NLRP3 inflammasome-dependent pyroptosis in macrophages facilitates liver fibrosis progression, and that the identified GSDMD may be used as a potential biomarker during liver fibrosis onset and progression.

Article Snippet: Detailed ELISA kits were as follows: Mouse S100A8 (E-EL-M3048, elabscience, Wuhan, Hubei, China), mouse IL-1β (VAL601, Novus, Littleton, CO, USA), mouse IL-18 (E-EL-M0730c, elabscience, Wuhan, Hubei, China), mouse GSDMD (JL46371-96T, JiangLai, Baoshan, Shanghai, China), human S100A8 (E-EL-H1289c, elabscience, Wuhan, Hubei, China), human IL-1β (Mengbio, Shapingba, Chongqing, China), human IL-18 (Mengbio, Shapingba, Chongqing, China), and human GSDMD (Mengbio, Shapingba, Chongqing, China).

Techniques: Biomarker Discovery

Journal: NPJ Vaccines

Article Title: Protective efficacy of the pan-fungal vaccine NXT-2 against vulvovaginal candidiasis in a murine model

doi: 10.1038/s41541-025-01171-4

Figure Lengend Snippet: IL-1β ( A ) and S100A8 ( B ) concentrations were assessed via ELISA. All data represents mean ± SEM. Statistical significance was assessed by using the Mann-Whitney test for IL-1β and S100A8 concentrations. C Tissue damage was assessed by measuring lactate dehydrogenase (LDH) activity in vaginal lavage fluid twenty-eight days post vaccination (28dpv) and five days post challenge (5dpc). Data represents mean ± SEM. Statistical significance was assessed by two-way ANOVA with Tukey correction (* P = 0.0111, *** P = 0.0007, **** P < 0.0001). D Quantification of inflammation in vaginal tissue from sham and NXT-2 immunized mice excised 5 days post challenge. Data represents mean ± SEM. Statistical significance was assessed by unpaired student t -test (**** P < 0.0001). Vaginal tissue excised following immunization and C. albicans challenge was stained with H&E (sham— E , NXT-2— G ) and the neutrophil specific marker anti-mouse Ly6G (sham— F , NXT-2— H ). Tissue sections were scanned at 40× objective magnification using the Aperio AT2 (Leica Biosystems). Image analysis and capture was performed using ImageScope (Leica Biosystems) at 20× digital zoom level.

Article Snippet: S100A8 : Utilizing the Mouse S100A8 ELISA kit (Rockland Immunochemicals), samples were diluted 1:100 and the manufacturer’s instructions were followed.

Techniques: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Activity Assay, Staining, Marker

Journal: Journal of Translational Medicine

Article Title: Single-cell RNA sequencing reveals recruitment of the M2-like CCL8 high macrophages in Lewis lung carcinoma-bearing mice following hypofractionated radiotherapy

doi: 10.1186/s12967-024-05118-6

Figure Lengend Snippet: Single-cell transcriptomic landscape of subcutaneous LLC tumors at early-stage following hypofractionated radiotherapy. A Overview of the experimental design for single-cell RNA sequencing. B Uniform Manifold Approximation (UMAP) plot showing the unsupervised clusters of 54883 single cells and annotated cell types. C The proportion of each cell types in LLC tumors treated with or without radiation. For B and C, each color represents the same cell type. D Feature plot and E Dot plot showing marker genes. F Heatmap showing serum chemokines level in LLC murine models 72 h post-treatment. Each row in the heatmap has been scaled. G The concentrations of CCL2, CCL7, CCL8, and CSF1 in serum measured by ELISA (n = 5/group). Data are presented as mean ± SD with student unpaired t test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns, not significant

Article Snippet: For proliferation assay and colony formation assay, the LLC cell line was treated with recombinant CCL8 (MedChemExpress, Cat#: HY-P7771) at 0, 5 ng/ml, 20 ng/ml.

Techniques: RNA Sequencing, Marker, Enzyme-linked Immunosorbent Assay

Journal: Journal of Translational Medicine

Article Title: Single-cell RNA sequencing reveals recruitment of the M2-like CCL8 high macrophages in Lewis lung carcinoma-bearing mice following hypofractionated radiotherapy

doi: 10.1186/s12967-024-05118-6

Figure Lengend Snippet: Identification of M2-like Ccl8 high Macrophages in the LLC-bearing murine model. A UMAP plot showing the annotation of macrophage populations in LLC tumors. B Feature plot and C Dotplot showing marker genes of macrophage populations. D Cell proportion of each cell type in NT and RT groups. E Heatmap displaying scores of M1, M2, angiogenesis, phagocytosis for each macrophage population. F Correlation analysis of CCL8 and CD163 expression level in TCGA-LUAD datasets (Spearman’s rho value = 0.542, p < 0.001). G The Gene Ontology (GO) enrichment analysis of each macrophage populations. H Heatmap displaying transcription factors activity of the Mac_Ccl8, Mac_Hmox1, Mono_Cxcl3, and Mono_Plac8 populations. I Development trajectory of macrophages populations predicted by Monocle2. J The cell density and K gene expression patterns along with the pseudotime,

Article Snippet: For proliferation assay and colony formation assay, the LLC cell line was treated with recombinant CCL8 (MedChemExpress, Cat#: HY-P7771) at 0, 5 ng/ml, 20 ng/ml.

Techniques: Marker, Expressing, Activity Assay, Gene Expression

Journal: Journal of Translational Medicine

Article Title: Single-cell RNA sequencing reveals recruitment of the M2-like CCL8 high macrophages in Lewis lung carcinoma-bearing mice following hypofractionated radiotherapy

doi: 10.1186/s12967-024-05118-6

Figure Lengend Snippet: Hypofractionated radiotherapy promoted the crosstalk between the Mac_Ccl8 and lymphocytes. A UMAP plot showing the annotation of lymphocyte populations. B Feature plot and C Dot plot showing marker genes of lymphocyte populations. D The proportion of each lymphocyte population in two groups. E Violin plots displaying the expression level of immune checkpoint ligand genes in each lymphocyte populations of NT and RT groups. F Circle plots showing number of interactions in two groups inferred by CellChat. G The comparison of cellular communication probability from Mac_Ccl8 to T and NK cells in between two groups. H Chord plot displaying the upregulated signaling pathways in the RT group relative to the NT group. I Heatmap of the differential interaction strength of the GALECTIN signaling pathway between two groups

Article Snippet: For proliferation assay and colony formation assay, the LLC cell line was treated with recombinant CCL8 (MedChemExpress, Cat#: HY-P7771) at 0, 5 ng/ml, 20 ng/ml.

Techniques: Marker, Expressing, Comparison, Protein-Protein interactions

Journal: Journal of Translational Medicine

Article Title: Single-cell RNA sequencing reveals recruitment of the M2-like CCL8 high macrophages in Lewis lung carcinoma-bearing mice following hypofractionated radiotherapy

doi: 10.1186/s12967-024-05118-6

Figure Lengend Snippet: Hypofractionated radiotherapy reprograms CCL8 high macrophages through the CCL signaling pathway. A Volcano plot showing differentially expressed genes of the Mac_Ccl8 between the NT and RT groups. Adjusted p value < 0.05, two-sided Wilcoxon test. B Violin plots comparing the expression of Ccl2, Ccl3, Ccl4, Ccl7, Ccl8, and Ccl12 in the NT and RT groups. Unpaired two-sided Wilcoxon test. C Representative examples of multiplex immunofluorescent labeling CD206 and CCL8. Green, CD206; Red, CCL8; Blue, DAPI. D Bar plots showing the GO enrichment analysis of upregulation and downregulation genes in the RT group. E Differences in IFN-Gamma and TNF pathways activity between two groups inferred by GSEA. F Cell–cell communication network between myeloid populations and LLC cells. G River plot displaying communication patterns of different cell types. H Chord plot (top) and heatmap (bottom) showing communication network of CCL signaling pathway in different cell types. I Weighted network analysis of differential interaction strength of signals in the Mac_Ccl8 population between two groups. J The comparison of cellular communication probability from Mac_Ccl8 to other myeloid populations between two groups. K Chord plot displaying the upregulated signaling pathways in the RT group relative to the NT group. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns, not significant

Article Snippet: For proliferation assay and colony formation assay, the LLC cell line was treated with recombinant CCL8 (MedChemExpress, Cat#: HY-P7771) at 0, 5 ng/ml, 20 ng/ml.

Techniques: Expressing, Multiplex Assay, Labeling, Activity Assay, Comparison, Protein-Protein interactions

Journal: Journal of Translational Medicine

Article Title: Single-cell RNA sequencing reveals recruitment of the M2-like CCL8 high macrophages in Lewis lung carcinoma-bearing mice following hypofractionated radiotherapy

doi: 10.1186/s12967-024-05118-6

Figure Lengend Snippet: Hypofractionated radiotherapy promotes M2-like Ccl8 high macrophages infiltration and leads to poor prognosis. A Kaplan–Meier plots showing worse clinical prognosis in the LUAD and HNSC patients with the higher expression level of the Mac_Ccl8 signature. HR, hazard ratio. B Schematic diagram of the combination treatment with hypofractionated radiotherapy and the Bindarit. The intraperitoneally administration of Bindarit began from day 5 to day 11 post-tumor injection, and radiation treatment was initiated from day 7 to day 9 post-tumor injection. C Growth curves of tumors in LLC-bearing mice in the indicated treatment groups (n = 5 mice/group). Data are presented as mean ± SD with two-way ANOVA test. D Representative examples in the indicated treatment groups of multiplex immunofluorescent labeling F4/80, CD206 and CCL8. Yellow, F4/80, Green, CD206; Red, CCL8; Blue, DAPI. E Percentages of M1 and M2 macrophages in the specific treatment groups analyzed by flow cytometry (n = 3/group). F Representative flow cytometry panels showing M2 macrophages (top) and M1 macrophages (bottom). BI, Bindarit; RT, Radiation therapy; RT + BI, the combination therapy of the radiation and the Bindarit

Article Snippet: For proliferation assay and colony formation assay, the LLC cell line was treated with recombinant CCL8 (MedChemExpress, Cat#: HY-P7771) at 0, 5 ng/ml, 20 ng/ml.

Techniques: Expressing, Injection, Multiplex Assay, Labeling, Flow Cytometry