Journal:

Article Title: Detection of Micrometastatic Tumor Cells in pN0 Lymph Nodes of Patients With Completely Resected Nonsmall Cell Lung Cancer

doi:

Figure Lengend Snippet: Figure 1. (A) A single micrometastatic cytokeratin-positive cell with deep brown-stained cytoplasm is seen against a background of benign lymphatic cells (original magnification ×400). (B) A single micrometastatic p53-positive cell with a deep brown-stained nucleus is seen against a background of benign lymphatic cells (original magnification ×400).

Article Snippet: The slides were stained with primary antibodies against the cytokeratin and p53 proteins by using a labeled streptavidin-biotin method (DAKO LSAB kit, Dako Corp., Carpinteria, CA).

Techniques: Staining

Journal:

Article Title: Detection of Micrometastatic Tumor Cells in pN0 Lymph Nodes of Patients With Completely Resected Nonsmall Cell Lung Cancer

doi:

Figure Lengend Snippet: Table 1. CLINICOPATHOLOGIC CHARACTERISTICS OF THE PATIENTS

Article Snippet: The slides were stained with primary antibodies against the cytokeratin and p53 proteins by using a labeled streptavidin-biotin method (DAKO LSAB kit, Dako Corp., Carpinteria, CA).

Techniques: Immunohistochemical staining, Immunohistochemistry

Journal:

Article Title: Detection of Micrometastatic Tumor Cells in pN0 Lymph Nodes of Patients With Completely Resected Nonsmall Cell Lung Cancer

doi:

Figure Lengend Snippet: Table 2. DISTRIBUTION OF PATIENTS WITH CK OR p53 PROTEIN-POSITIVE CELLS

Article Snippet: The slides were stained with primary antibodies against the cytokeratin and p53 proteins by using a labeled streptavidin-biotin method (DAKO LSAB kit, Dako Corp., Carpinteria, CA).

Techniques:

Journal:

Article Title: Detection of Micrometastatic Tumor Cells in pN0 Lymph Nodes of Patients With Completely Resected Nonsmall Cell Lung Cancer

doi:

Figure Lengend Snippet: Table 3. RECURRENCE RATES

Article Snippet: The slides were stained with primary antibodies against the cytokeratin and p53 proteins by using a labeled streptavidin-biotin method (DAKO LSAB kit, Dako Corp., Carpinteria, CA).

Techniques: Immunohistochemical staining

Journal:

Article Title: Detection of Micrometastatic Tumor Cells in pN0 Lymph Nodes of Patients With Completely Resected Nonsmall Cell Lung Cancer

doi:

Figure Lengend Snippet: Figure 2. (A) Survival of patients with stage 1 disease (n = 49) with or without micrometastatic tumor cells in the pN0 lymph nodes detected by the combination of cytokeratin and p53 protein immunohistochemical staining (Kaplan-Meier analysis, log-rank test, P = .0003). (B) Survival of patients with stage 1A disease (n = 18) with or without micrometastatic tumor cells in the pN0 lymph nodes detected by the combination of cytokeratin and p53 protein immunohistochemical staining (P = .057). (C) Survival of patients with stage 1B disease (n = 31) with or without micrometastatic tumor cells in the pN0 lymph nodes detected by the combination of cytokeratin and p53 protein immunohistochemical staining (P = .0016). “CK/p53 positive” included cytokeratin-positive/p53-positive, cytokeratin-positive/p53-negative, and cytokeratin-negative/p53-positive immunohistochemical stainings. “CK/p53 negative” included cytokeratin-negative/p53-negative immuno-histochemical staining.

Article Snippet: The slides were stained with primary antibodies against the cytokeratin and p53 proteins by using a labeled streptavidin-biotin method (DAKO LSAB kit, Dako Corp., Carpinteria, CA).

Techniques: Immunohistochemical staining, Staining

Journal:

Article Title: Detection of Micrometastatic Tumor Cells in pN0 Lymph Nodes of Patients With Completely Resected Nonsmall Cell Lung Cancer

doi:

Figure Lengend Snippet: Table 4. MULTIVARIATE COX REGRESSION ANALYSIS OF PROGNOSTIC FACTORS

Article Snippet: The slides were stained with primary antibodies against the cytokeratin and p53 proteins by using a labeled streptavidin-biotin method (DAKO LSAB kit, Dako Corp., Carpinteria, CA).

Techniques: Immunohistochemical staining

Journal: Nucleic acids research

Article Title: Nuclear AGO2 supports influenza A virus replication through type-I interferon regulation.

doi: 10.1093/nar/gkaf268

Figure Lengend Snippet: Figure 2. p53 associates with AGO2 in the nucleus ( A ) STRING protein-protein interaction network of AGO2. ( B ) Representative AGO2 and p53 immunoblots from cytoplasmic (C) and nuclear (N) lysates in HEK293 cells treated with Doxorubicin (Doxo) for 24 h. GAPDH served as a cytoplasmic marker and HIST3H3 served as nuclear marker. n = 3 ( C ) AGO2 IP from HEK293 cells treated with Doxorubicin (Doxo) for 24 h. Representative immunoblots of AGO2 and p53. n = 3 ( D ) R epresentativ e AGO2, p53 and SV40 immunoblots from cytoplasmic (C) and nuclear (N) lysates in HEK293 cells transfected with SV40 large T antigen expressing plasmid. GAPDH served as a cytoplasmic marker and HIST3H3 served as nuclear marker. n = 3 ( E ) same as in (D) e x cept f or immunoblots f or NS1 in HEK293 cells transfected with NS1 mCherry e xpressing plasmid f or 24 h. n = 3 ( F ) R epresentativ e AGO2 and p53 immunoblots from cytoplasmic (C) and nuclear (N) lysates in HEK293 cells infected with PR8 virus at MOI 10 for 16 h. GAPDH served as cytoplasmic marker and HIST3H3 as nuclear marker. n = 3 ( G ) AGO2 IP from HEK293 cells treated with PR8 virus at MOI 10 for 16 h. Representative immunoblots of AGO2 and p53. n = 3 ( H ) AGO2 IP from cytoplasmic and nuclear fractions in HEK293 cells transfected with SV40 large T antigen expressing plasmid for 24 h. Representative immunoblots of AGO2 and p53. n = 3 ( I ) same as in (H) except for HEK293 cells were transfected with WT NS1 expressing plasmid for 24 h. n = 3.

Article Snippet: The pull-down efficiency was confirmed by western blot. p53 mutagenesis The plasmid encoding amino acids 1–393 of the p53 protein with a FLAG tag at the C-terminus (Addgene plasmid #10 838) was used as a template for site-directed mutagenesis to delete the N -terminal regions spanning amino acids 1– 31, 1–62, or 1–93.

Techniques: Western Blot, Marker, Transfection, Expressing, Plasmid Preparation, Infection, Virus

Journal: Nucleic acids research

Article Title: Nuclear AGO2 supports influenza A virus replication through type-I interferon regulation.

doi: 10.1093/nar/gkaf268

Figure Lengend Snippet: Figure 3. Tetrameric p53 protects nuclear AGO2 from proteasomal degradation ( A ) Schematic diagram of human p53 protein and N -terminal truncated Flag-tagged p53 isoforms used in the study. ( B ) Representative Flag immunoblots from HEK293 cells transfected with Flag-WT-p53 or N -terminally Flag-t agged p53 mut ants. G APDH serv ed as a loading control. n = 3 ( C ) Flag IP from HEK293 cells transfected with plasmids expressing Flag-WT-p53 or N -terminally Flag-tagged p53 mutant. HEK293 cells were transfected with the p53 plasmids for 1 day and then treated with 1 μg / mL Do x orubicin (Do x o) for 1 more day. Representative immunoblots of AGO2 and Flag are indicated. n = 3 ( D ) Representative AGO2 and p53 immunoblots from cytoplasmic (C) and nuclear (N) lysates in A549 cells treated with 0.5 μg / mL arsenic trioxide (ATO) for 24 h. GAPDH served as a cytoplasmic marker and HIST3H3 served as nuclear marker. n = 3 ( E ) p53 IP from A549 cells treated with 0.5 μg / mL Arsenite trioxide (ATO) for 24 h. Representative immunoblots of AGO2 and p53. n = 3 ( F ) Docking model of the PIWI domain of AGO2 with the N -terminal region of p53 was generated using the HDOCK server. The PIWI domain is depicted in green, and the N -terminal region of p53 is shown in pink (model 4) in ribbon representation. ( G ) AGO2 IP from cytoplasmic (C) and nuclear (N) lysates in A549 cells. Representative immunoblots of AGO2, p53 and TNRC6A. n = 3 ( H ) p53 IP from cytoplasmic (C) and nuclear (N) lysates in A549 cells. Representative immunoblots of AGO2, p53 and TNRC6A. n = 3 ( I ) TNRC6 IP from cytoplasmic (C) and nuclear (N) lysates in A549 cells treated with siRNAs for 24 h. siCntr: control scramble siRNAs; and siAGO2: siRNA specific for AGO2. R epresentativ e TNR C6, AGO2 and p53 immunoblots. GAPDH served as a cytoplasmic marker and HIST3H3 served as nuclear marker. ( J ) Representative AGO2 and p53 immunoblots from cytoplasmic (C) and nuclear (N) lysates in A549 cells treated with siRNAs for 24 h. siCntr: control scramble siRNAs; siTP53: siRNA specific for TP53; and siAGO2: two different siRNAs specific for AGO2. GAPDH served as a cytoplasmic marker and HIST3H3 served as nuclear marker. n = 3.

Article Snippet: The pull-down efficiency was confirmed by western blot. p53 mutagenesis The plasmid encoding amino acids 1–393 of the p53 protein with a FLAG tag at the C-terminus (Addgene plasmid #10 838) was used as a template for site-directed mutagenesis to delete the N -terminal regions spanning amino acids 1– 31, 1–62, or 1–93.

Techniques: Western Blot, Transfection, Control, Expressing, Mutagenesis, Marker, Generated

Journal: Nucleic acids research

Article Title: Nuclear AGO2 supports influenza A virus replication through type-I interferon regulation.

doi: 10.1093/nar/gkaf268

Figure Lengend Snippet: Figure 6. Nuclear AGO2 downregulates type-I IFN pathway genes and TRIM71 in IA V -infected cells ( A ) Distribution of fPAR-CLIP sequence reads in clusters across target RNA across 3 ′ UTR, coding sequence, 5 ′ UTR, and introns from cytoplasmic (C) and nuclear (N) fractions of HEK293 cells infected with PR8 virus at MOI 10 for 16 h. The graph shows the average of two independent experiments. ( B ) Scatter plot of cytoplasmic fPAR-CLIP log2 normalized signal in control (- Virus) and IAV infected (+Virus) HEK293 cells. Each dots represents a gene and is the a v erage of two independent experiments. Blue dashed line shows a perfect correlation. Black dots are genes with higher PAR-CLIP signal in the IA V -infected sample while gray dots are genes with higher PAR-CLIP signal in the control sample. ( C ) same as in (B) but for nuclear fPAR-CLIP. ( D ) CDF of AGO1-4 targets in control cytoplasmic fraction compared to non-targets. Targets are the genes depicted as grey dots in (B) ( E ) CDF of AGO1-4 targets in virus infected cytoplasmic fraction compared to non-targets. Targets are the genes depicted as black dots in (B) ( F ) CDF of AGO1-4 targets in control nuclear fraction compared to non-targets. Targets are the genes depicted as gray dots in (C) ( G ) CDF of AGO1-4 targets in virus infected nuclear fraction compared to non-targets. Targets are the genes depicted as black dots in (C) ( H ) same graph as in (C) but with type-I IFN pathw a y genes highlighted in golden y ello w. ( I ) same as in (G) but for type-I IFN pathway genes. Axis is cut at +2 log 2 fold changes for visualization purposes. ( J ) Rank plot showing AGO targets, ordered by normalized PAR-CLIP signal, in IAV infected, nuclear fraction of HEK293 cells. Each dot represents a gene and is the average of two independent experiments. ( K ) TRIM71 IGV track in nuclear fraction of in control (- Virus) and IAV infected (+Virus) HEK293 cells. ( L ) R epresentativ e TRIM71 immunoblots from whole cell lysates in HEK293 infected with PR8 virus at MOI 10 for 16 h. GAPDH served as a loading control. n = 3 ( M ) R epresentativ e HA-TRIM71, AGO2 and p53 immunoblots from cytoplasmic (C) and nuclear (N) lysates in HEK293 cells transfected with HA-TRIM71 and infected with PR8 virus at MOI 10 for 16 h. GAPDH served as a cytoplasmic marker and HIST3H3 served as nuclear marker. n = 3, ( N ) Relative expression, as measured by RT-qPCR, of IFNB mRNA levels in HEK293 cells transfected with HA-TRIM71 and infected with PR8 virus at MOI 10 for 16 hours. GAPDH was used as a reference gene. Bars are mean and error bars represent ± SD. ∗∗P < 0.01 by unpaired t-test. n = 3 ( O ) Relative expression, as measured by RT-qPCR, of HA and NS1 mRNA levels in HEK293 cells transfected with HA-TRIM71 and infected with PR8 virus at MOI 10 for 16 h. GAPDH was used as a reference gene. Bars are mean and error bars represent ± SD. ∗P < 0.05, ∗∗P < 0.01 by unpaired t-test. n = 3.

Article Snippet: The pull-down efficiency was confirmed by western blot. p53 mutagenesis The plasmid encoding amino acids 1–393 of the p53 protein with a FLAG tag at the C-terminus (Addgene plasmid #10 838) was used as a template for site-directed mutagenesis to delete the N -terminal regions spanning amino acids 1– 31, 1–62, or 1–93.

Techniques: Infection, Sequencing, Virus, Control, Western Blot, Transfection, Marker, Expressing, Quantitative RT-PCR