Journal: Clinical Cancer Research

Article Title: Overexpression of PD-L1 Significantly Associates with Tumor Aggressiveness and Postoperative Recurrence in Human Hepatocellular Carcinoma

doi: 10.1158/1078-0432.ccr-08-1608

Figure Lengend Snippet: Fig. 1. PD-Ls expression in HCC cell lines and tissue samples.Western blots detected constitutive PD-L1and PD-L2 expressions on 7 HCC cell lines (A). Total protein (50 Ag) was loaded per lane. Consecutive sections were used for immunohistochemical study on expression of PD-L1and PD-L2 on HCC tumor tissues. Representative cases of concurrent high or low (B) as well as either high (C) expression of PD-L1and PD-L2 on HCC tumor tissues were shown. Positive cells were stained brown. Magnification, 200.

Article Snippet: The cells lysing, protein extraction, concentration detection, and Western analysis were done as we described previously (31, 34) with the following primary antibodies: goat antihuman PD-L1 (1:1,000) and PD-L2 (1:1,000) monoclonal antibodies (R&D Systems) with h-actin (1:500; Chemicon) as an internal loading control.

Techniques: Expressing, Western Blot, Immunohistochemical staining, Staining

Journal: Clinical Cancer Research

Article Title: Overexpression of PD-L1 Significantly Associates with Tumor Aggressiveness and Postoperative Recurrence in Human Hepatocellular Carcinoma

doi: 10.1158/1078-0432.ccr-08-1608

Figure Lengend Snippet: Fig. 2. Kaplan-Meier curves of survival differences among HCC patients. DFS and OS for expression of PD-L1 (A), PD-L2 (B), and their combination (C) were found to be statistically significant. Significant differences in DFS and OS were validated in an independent cohort based on PD-L1 expression (D). P values were determined by the log-rank test.

Article Snippet: The cells lysing, protein extraction, concentration detection, and Western analysis were done as we described previously (31, 34) with the following primary antibodies: goat antihuman PD-L1 (1:1,000) and PD-L2 (1:1,000) monoclonal antibodies (R&D Systems) with h-actin (1:500; Chemicon) as an internal loading control.

Techniques: Expressing

Journal: Retrovirology

Article Title: Increased BST2 expression during simian immunodeficiency virus infection is not a determinant of disease progression in rhesus monkeys

doi: 10.1186/s12977-015-0219-8

Figure Lengend Snippet: BST2 mRNA induction by type I interferon. a Fold induction of relative BST2 mRNA in PBMC from three uninfected rhesus macaques after stimulation with human Interferon Alpha A (Alpha 2a) for 16 h. Data are expressed as fold increase over baseline after normalization to pre-treatment values. Error bars represent standard deviation, b relative mRNA copies of BST2 in PBMC (shown in copy numbers per 100 copies of GAPDH) are illustrated in relation to plasma IFN-alpha levels from blood samples of 18 uninfected rhesus macaques 24 h after inoculation of replication incompetent adenovirus or fowl pox vectors. The black dashed line indicates the detection limit of the ELISA and c whole blood MX1 mRNA levels correlate with BST2 mRNA determined in 38 SIVmac251 infected rhesus macaques at 24 wpi. Relative mRNA levels are depicted as log-transformed copy numbers per 100 copies of GAPDH. Each data point represents one animal. Regression line is shown; r , Spearman’s correlation coefficient; p , p value

Article Snippet: PBMC from three healthy animals were stimulated with 10, 50 and 100 ng Human Interferon Alpha A (Alpha 2a) (PBL Biomedical Laboratories) for 16 h. Fold induction of BST2 mRNA expression was calculated using GAPDH as internal control.

Techniques: Standard Deviation, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Infection, Transformation Assay

Journal: Cell Reports Methods

Article Title: Generation, expansion, gene delivery, and single-cell profiling in rhesus macaque plasma B cells

doi: 10.1016/j.crmeth.2024.100878

Figure Lengend Snippet:

Article Snippet: IFN alpha 2B , Proteintech , HZ-1072.

Techniques: Enzyme-linked Immunospot, Isolation, Enzyme-linked Immunosorbent Assay, Recombinant, Software

Journal: Cell reports

Article Title: A Human Papillomavirus-Independent Cervical Cancer Animal Model Reveals Unconventional Mechanisms of Cervical Carcinogenesis

doi: 10.1016/j.celrep.2019.02.004

Figure Lengend Snippet: (A) Quantitative experiments showing mRNA levels of major components of the IFNRs/JAK/STAT/IRF9 pathway and downstream target genes in control (hCerEC-Mx) and hCerEC-YAP S127A cells. Each bar represents the mean ± SEM (n = 4). *p < 0.05, **p < 0.01, and ***p < 0.001. (B) Quantitative data showing mRNA levels of YAP1 and genes of the core components of the IFNR/JAK/STAT antiviral pathway in hCerEC-CTRL cells (cells transfected with scrambled non-target siRNA) and hCerEC-siYAP cells (cells transfected with YAP1-specific siRNA). Each bar represents the mean ± SEM (n = 4). *p < 0.05, **p < 0.01, and ***p < 0.001. (C) Representative blots from three independent experiments showing expression and activation of key proteins and kinases in the JAK/STAT/IRF9 pathway in hCerEC-MX (control), hCerEC-YAP, and hCerEC-YAP S127A cells. (D) Representative images showing HPV16 PsV-derived GFP signal in hCerEC-MX, hCerEC-YAP, and hCerEC-YAP S127A cells in the presence or absence of recombined human interferon alpha 2b (IFNα2b). GFP signal indicates the infection efficiency of HPV16 PsV in these cells. Scale bar: 100 μm. Quantitative results of GFP signal is presented in . (E) RT-PCR analyses showing expression of JAK1 , IRF9 , STAT1 , and MX1 mRNA in cervical tissues from control KRT14-rtTA (CTRL) and KRT14-YAP S127A mice. Each bar represents the mean + SEM (n = 4). ***p < 0.001. (F) Representative images showing expression of JAK1, IRF9, MX1, and ISG15 proteins in cervical tissues from KRT14-rtTA control (CTRL) and KRT14-YAP S127A mice. Protein expression was determined using the peroxidase-based immunohistochemistry. The nuclei were counterstained with hematoxylin. Scale bar: 250 μm.

Article Snippet: Interferon alpha 2b (IFNα2b, #11105–1) was purchased from R&D systems Inc. (Minneapolis, MN).

Techniques: Control, Transfection, Expressing, Activation Assay, Derivative Assay, Infection, Reverse Transcription Polymerase Chain Reaction, Immunohistochemistry

Journal: Cell reports

Article Title: A Human Papillomavirus-Independent Cervical Cancer Animal Model Reveals Unconventional Mechanisms of Cervical Carcinogenesis

doi: 10.1016/j.celrep.2019.02.004

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Interferon alpha 2b (IFNα2b, #11105–1) was purchased from R&D systems Inc. (Minneapolis, MN).

Techniques: Virus, Microarray, Recombinant, Reverse Transcription, SYBR Green Assay, Plasmid Preparation, Staining, Blocking Assay

Journal: Nucleic acids research

Article Title: Internal RNA 2'O-methylation in the HIV-1 genome counteracts ISG20 nuclease-mediated antiviral effect.

doi: 10.1093/nar/gkac996

Figure Lengend Snippet: Figure 7. Restriction profile of ISG20 WT and the R53A/D90A mutant on replication competent HIV-1. (A) HeLa P4C5 cells were treated or not with 1000 IU/ml human IFN-2a for 18h and transfected with a siRNA control (−) or targeting ISG20 (+), as indicated. At 48h post-transfection, ISG20 expression was assessed by western-blot using anti-ISG20 antibodies. (B–D) HeLa P4C5 cells were treated or not with 1000 IU/ml human IFN-I for 18 h and transfected with a siRNA control (−) or targeting ISG20 (+), as indicated. At 48h post-transfection, cells were infected with HIV-1 NL4-3 produced in WT or in FTSJ3-KO cells. At 6 h post-infection, cells were harvested and total RNA and DNA were extracted for specific qPCR analysis. (B) The expression of endogenous ISG20 upon IFN-I induction was evaluated by RT-qPCR. (C) Quantification of ectopic expression of transfected ISG20 WT and double mutant by RT-qPCR. (D) The amount of HIV-1 RT products was estimated by qPCR. Data represent the mean of three independent experiments performed in duplicate ± SD. ****P < 0.0001, as determined by one-way ANOVA with Bonferroni post hoc test. ns, non-significant.

Article Snippet: When indicated, HeLa P4C5 and Jurkat cells were treated with recombinant human IFN- 2a (R&D Systems).

Techniques: Mutagenesis, Transfection, Control, Expressing, Western Blot, Infection, Produced, Quantitative RT-PCR