Journal: Cancer Management and Research

Article Title: Tumor-Associated CD163 + M2 Macrophage Infiltration is Highly Associated with PD-L1 Expression in Cervical Cancer

doi: 10.2147/CMAR.S257692

Figure Lengend Snippet: Immunohistochemistry staining for PD-L1, CD68, and CD163 expression in cervical squamous cell carcinoma tissues. The left panel shows negative staining for PD-L1 and low macrophage density. The right panel shows positive staining for PD-L1 and high macrophage density.

Article Snippet: After placing in a microwave oven for pretreatment (PD-L1: EDTA antigen repair solution for 30 min, pH 9; CD68 and CD163: sodium citrate buffer for 15 min, pH 6), endogenous peroxidase was inhibited by 3% H 2 O 2 for 30 min, then the sections were incubated with 10% normal goat serum for 40 min. Primary antibodies composed of rabbit anti-PD-L1 antibody 28–8 (diluted 1:100, ab205921, Abcam, Cambridge, UK), rabbi anti-CD68 antibody (diluted 1:50, BA3638, Boster, California, UK), or rabbit anti-CD163 antibody (diluted 1:200, bs-2527R, bioss, Beijing, China), were applied overnight in a moist chamber at 4°C.

Techniques: Immunohistochemistry, Staining, Expressing, Negative Staining

Journal: Cancer Management and Research

Article Title: Tumor-Associated CD163 + M2 Macrophage Infiltration is Highly Associated with PD-L1 Expression in Cervical Cancer

doi: 10.2147/CMAR.S257692

Figure Lengend Snippet: Density of CD68- and CD163-Positive Cells in the Tumor Field and Clinical Features in 120 Samples of Cervical Squamous Cell Carcinoma

Article Snippet: After placing in a microwave oven for pretreatment (PD-L1: EDTA antigen repair solution for 30 min, pH 9; CD68 and CD163: sodium citrate buffer for 15 min, pH 6), endogenous peroxidase was inhibited by 3% H 2 O 2 for 30 min, then the sections were incubated with 10% normal goat serum for 40 min. Primary antibodies composed of rabbit anti-PD-L1 antibody 28–8 (diluted 1:100, ab205921, Abcam, Cambridge, UK), rabbi anti-CD68 antibody (diluted 1:50, BA3638, Boster, California, UK), or rabbit anti-CD163 antibody (diluted 1:200, bs-2527R, bioss, Beijing, China), were applied overnight in a moist chamber at 4°C.

Techniques: Biomarker Discovery

Journal: Cancer Management and Research

Article Title: Tumor-Associated CD163 + M2 Macrophage Infiltration is Highly Associated with PD-L1 Expression in Cervical Cancer

doi: 10.2147/CMAR.S257692

Figure Lengend Snippet: Correlation of PD-L1 expression and density of CD68-and CD163-positive cells in cervical cancer. Notes: ( A ) PD-L1 expression in tumor cells and density ofCD68. ( B ) PD-L1 expression in tumor cells and density of CD163. ( C ) PD-L1 expression in the stroma and density of CD68. ( D ) PD-L1 expression in the stroma and density of CD163.

Article Snippet: After placing in a microwave oven for pretreatment (PD-L1: EDTA antigen repair solution for 30 min, pH 9; CD68 and CD163: sodium citrate buffer for 15 min, pH 6), endogenous peroxidase was inhibited by 3% H 2 O 2 for 30 min, then the sections were incubated with 10% normal goat serum for 40 min. Primary antibodies composed of rabbit anti-PD-L1 antibody 28–8 (diluted 1:100, ab205921, Abcam, Cambridge, UK), rabbi anti-CD68 antibody (diluted 1:50, BA3638, Boster, California, UK), or rabbit anti-CD163 antibody (diluted 1:200, bs-2527R, bioss, Beijing, China), were applied overnight in a moist chamber at 4°C.

Techniques: Expressing

Journal: Cancer Management and Research

Article Title: Tumor-Associated CD163 + M2 Macrophage Infiltration is Highly Associated with PD-L1 Expression in Cervical Cancer

doi: 10.2147/CMAR.S257692

Figure Lengend Snippet: Relationship between the density of CD68- and CD163-positive cells and the expression of PD-L1 in cervical cancer. ( A ) Density of CD68 in PD-L1 positive and negative expression groups in TC. ( B ) Density of CD163 in PD-L1 positive and negative expression groups in TC. ( C ) Density of CD68 in PD-L1 positive and negative expression groups in stroma. ( D ) Density of CD163 in PD-L1 positive and negative expression groups in stroma.

Article Snippet: After placing in a microwave oven for pretreatment (PD-L1: EDTA antigen repair solution for 30 min, pH 9; CD68 and CD163: sodium citrate buffer for 15 min, pH 6), endogenous peroxidase was inhibited by 3% H 2 O 2 for 30 min, then the sections were incubated with 10% normal goat serum for 40 min. Primary antibodies composed of rabbit anti-PD-L1 antibody 28–8 (diluted 1:100, ab205921, Abcam, Cambridge, UK), rabbi anti-CD68 antibody (diluted 1:50, BA3638, Boster, California, UK), or rabbit anti-CD163 antibody (diluted 1:200, bs-2527R, bioss, Beijing, China), were applied overnight in a moist chamber at 4°C.

Techniques: Expressing

Journal: Cancer Management and Research

Article Title: Tumor-Associated CD163 + M2 Macrophage Infiltration is Highly Associated with PD-L1 Expression in Cervical Cancer

doi: 10.2147/CMAR.S257692

Figure Lengend Snippet: Univariate and Multivariate Analysis of Factors Associated with PD-L1 Expression in Tumor Cells

Article Snippet: After placing in a microwave oven for pretreatment (PD-L1: EDTA antigen repair solution for 30 min, pH 9; CD68 and CD163: sodium citrate buffer for 15 min, pH 6), endogenous peroxidase was inhibited by 3% H 2 O 2 for 30 min, then the sections were incubated with 10% normal goat serum for 40 min. Primary antibodies composed of rabbit anti-PD-L1 antibody 28–8 (diluted 1:100, ab205921, Abcam, Cambridge, UK), rabbi anti-CD68 antibody (diluted 1:50, BA3638, Boster, California, UK), or rabbit anti-CD163 antibody (diluted 1:200, bs-2527R, bioss, Beijing, China), were applied overnight in a moist chamber at 4°C.

Techniques: Expressing, Biomarker Discovery

Journal: Cancer Management and Research

Article Title: Tumor-Associated CD163 + M2 Macrophage Infiltration is Highly Associated with PD-L1 Expression in Cervical Cancer

doi: 10.2147/CMAR.S257692

Figure Lengend Snippet: Recurrence-free and overall survival curves in CC patients. ( A ) Kaplan–Meier OS curves according to PD-L1 expression in tumor cells ( B ) Kaplan–Meier OS curves according to PD-L1 expression in stroma ( C ) Kaplan–Meier OS curves according to the density of CD68-positive cells ( D ) Kaplan–Meier OS curves according to the density of CD163-positive cells ( E ) Kaplan–Meier RFS curves according to PD-L1 expression in tumor cells ( F ) Kaplan–Meier RFS curves according to PD-L1 expression in stroma ( G ) Kaplan–Meier RFS curves according to the density of CD68-positive cells ( H ) Kaplan–Meier RFS curves according to the density of CD163-positive cells. (The density of CD68/CD163 above median = high CD68/CD163; The density of CD68/CD163 below median = low CD68/CD163; p -values obtained from Log-rank tests.).

Article Snippet: After placing in a microwave oven for pretreatment (PD-L1: EDTA antigen repair solution for 30 min, pH 9; CD68 and CD163: sodium citrate buffer for 15 min, pH 6), endogenous peroxidase was inhibited by 3% H 2 O 2 for 30 min, then the sections were incubated with 10% normal goat serum for 40 min. Primary antibodies composed of rabbit anti-PD-L1 antibody 28–8 (diluted 1:100, ab205921, Abcam, Cambridge, UK), rabbi anti-CD68 antibody (diluted 1:50, BA3638, Boster, California, UK), or rabbit anti-CD163 antibody (diluted 1:200, bs-2527R, bioss, Beijing, China), were applied overnight in a moist chamber at 4°C.

Techniques: Expressing

Journal: Cancer Management and Research

Article Title: Tumor-Associated CD163 + M2 Macrophage Infiltration is Highly Associated with PD-L1 Expression in Cervical Cancer

doi: 10.2147/CMAR.S257692

Figure Lengend Snippet: Univariate Analysis of Overall and Recurrence-Free Survival

Article Snippet: After placing in a microwave oven for pretreatment (PD-L1: EDTA antigen repair solution for 30 min, pH 9; CD68 and CD163: sodium citrate buffer for 15 min, pH 6), endogenous peroxidase was inhibited by 3% H 2 O 2 for 30 min, then the sections were incubated with 10% normal goat serum for 40 min. Primary antibodies composed of rabbit anti-PD-L1 antibody 28–8 (diluted 1:100, ab205921, Abcam, Cambridge, UK), rabbi anti-CD68 antibody (diluted 1:50, BA3638, Boster, California, UK), or rabbit anti-CD163 antibody (diluted 1:200, bs-2527R, bioss, Beijing, China), were applied overnight in a moist chamber at 4°C.

Techniques: Expressing

Journal: Experimental eye research

Article Title: A modified high-yield method for primary culture of rat retinal microglial cells.

doi: 10.1016/j.exer.2021.108919

Figure Lengend Snippet: Fig. 4. Primary rat retinal microglial cells were activated by LPS stimulation. (A) Microglial cells were treated with LPS (100 ng/mL) for 8 h on day 2. Phase-contrast microscopy images showed an abrupt change in morphology from ramified state (red arrow) to amoeboid (red arrowhead) after LPS stimulation; Scale bar: 100 μm; (B) Representative immunofluorescent images of primary retinal microglia stained with CD68 (red), and DAPI (blue). The expression of CD68 was upregulated after LPS (100 ng/mL) treatment for 8h; Scale bar: 50 μm. (C) Representative immunofluorescent images of primary retinal microglia with fluorescent beads (green) and nuclei labeled with DAPI (blue); microglial cells with phagocytosed beads were increased after LPS stimulation for 4h. The experiment was performed three times. Scale bar: 50 μm. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The primary antibodies used in this study were rabbit anti-IBA1 antibody (Abcam, ab178847, 1:500), rabbit anti-CD68 antibody (Boster, BA3638, 1:200), rat anti-CD11b antibody (Abcam, ab8878, 1:500), rabbit anti-GFAP antibody (Abcam, ab7260, 1:200), and rabbit antiTMEM119 antibody (Proteintech, 27585-1-AP, 1:200); the nuclei were stained with DAPI (Beyotime Biotechnology, C1002).

Techniques: Microscopy, Staining, Expressing, Labeling