Journal: Experimental Hematology & Oncology

Article Title: ScRNA-seq revealed an immunosuppression state and tumor microenvironment heterogeneity related to lymph node metastasis in prostate cancer

doi: 10.1186/s40164-023-00407-0

Figure Lengend Snippet: Primer sequences used in qRT-PCR

Article Snippet: They were then fixed with 4% formaldehyde (Biyuntian, P0099) at room temperature for 1 h, washed with PBS three times, permeabilized with 2% Triton X-100 (Biyuntian, P0096) for 5–10 min, washed with PBS three times, and blocked with 5% BSA at room temperature for 1 h. Primary antibodies for MYC (Thermo Fisher, MA1-980, 1:100), PD-L1 (FineTest, FNab06280, 1:50), CD47 (Bioss, bs-21460R, 1:100), CCL5 (Thermo Fisher, 710001,1:250), and EEF1A2 (Abcam, ab212172, 1:100) were diluted with 1% BSA, added to the samples, and incubated overnight at 4 °C in a humidified chamber (and reheated for 30 min the next day).

Techniques:

Journal: Experimental Hematology & Oncology

Article Title: ScRNA-seq revealed an immunosuppression state and tumor microenvironment heterogeneity related to lymph node metastasis in prostate cancer

doi: 10.1186/s40164-023-00407-0

Figure Lengend Snippet: MYC drives tumor progression through CD47 and PD-L1. ( A ). Immunohistochemistry analysis of MYC through PCa tissue microarray; N: Normal prostate tissue, T: Prostate cancer tissue, LM: lymphatic metastases; ( B ). Immunofluorescence of MYC, PDL1, and CD47 in LNCap; ( C ). Bioinformatics analysis of binding sites of MYC to PDL1 and CD47; ( D ). MYCmRNA, PDL1mRNA, and CD47mRNA expression in LNCaP after MYC down-regulation. ( E–F ). MYC, PDL1, and CD47 western blot protein expression analysis in LNCaP; ( G–H ). Metastatic ability of LNCAP cells analyzed using transwell assay after down-regulation of MYC.

Article Snippet: They were then fixed with 4% formaldehyde (Biyuntian, P0099) at room temperature for 1 h, washed with PBS three times, permeabilized with 2% Triton X-100 (Biyuntian, P0096) for 5–10 min, washed with PBS three times, and blocked with 5% BSA at room temperature for 1 h. Primary antibodies for MYC (Thermo Fisher, MA1-980, 1:100), PD-L1 (FineTest, FNab06280, 1:50), CD47 (Bioss, bs-21460R, 1:100), CCL5 (Thermo Fisher, 710001,1:250), and EEF1A2 (Abcam, ab212172, 1:100) were diluted with 1% BSA, added to the samples, and incubated overnight at 4 °C in a humidified chamber (and reheated for 30 min the next day).

Techniques: Immunohistochemistry, Microarray, Immunofluorescence, Binding Assay, Expressing, Western Blot, Transwell Assay