Journal: Science advances

Article Title: Avian ANP32A incorporated in avian influenza A virions promotes interspecies transmission by priming early viral replication in mammals.

doi: 10.1126/sciadv.adj4163

Figure Lengend Snippet: Fig. 7. avANP32A transferred by avian influenza A virus accelerates the process of obtaining adaptive mutations. (A) H9N2 virus produced from HEK293T cells overexpressing Flag-tagged ANP32A or with an empty vector were purified by ultracentrifugation with a prior HAd step. The cell lysates and purified virions were then subjected to Western blotting. (B) Viral replication in MDCK cells or MDCK-avANP32A cells infected with H9N2 (huANP32A) virus and H9N2 (avANP32A) virus at an MOI of 0.1. Error bars represent mean ± SD from n = 3 independent biological replicates; unpaired t test; **P < 0.01 and ****P < 0.0001. (C) Model for the effect of avANP32A transferred by avian influenza A virus on viral replication and adaptive mutation acquisition when jumping from avian hosts to mammalian hosts. (D) H9N2 virus pack- aged with either avANP32A or huANP32A was blind passaged six times in MDCK cells. Viral RNAs were extracted and the C terminus of PB2 was amplified and deep- sequenced to monitor the residue phenotype of PB2 627 and 701 positions during passages in MDCK cells. Bar graph represents the percentage of adaptive mutations including PB2-E627K/V and D701N in each passage. Error bars represent mean ± SEM from n = 4 independent biological replicates, unpaired t test; *P < 0.05. (E) Sche- matic representation of the protocol for the experiments shown in (F). (F) The lungs of infected mice were collected for viral RNA extraction, and the C terminus of PB2 was amplified and cloned into T vectors. Nine molecular clones from each sample were randomly picked and sequenced for determination of the residue phenotype of PB2 627 and 701 positions. Samples with nine clone negatives for the emergence of PB2-E627K/V and D701N were recognized as no occurrence of adaptive mutations. In each group, the percentage of mice with emergence of adaptive mutations was calculated.

Article Snippet: Immunoblotting analysis was carried out using the following primary antibodies: ANP32A rabbit polyclonal (15810- 1- AP, Proteintech), ANP32B mouse monoclonal (66160- 1- Ig, Proteintech), ANP32E rabbit polyclonal (A17220, Abclonal), Flag mouse monoclonal (F1804, SigmaAldrich), Flag rabbit polyclonal (F7425, Sigma- Aldrich), HA mouse monoclonal (H9658, Sigma- Aldrich), ACTB rabbit monoclonal (AC026, Abclonal), His mouse monoclonal (66005- 1, Proteintech), V5 mouse monoclonal (ab27671, Abcam), glutathione S- transferase (GST) rabbit polyclonal (10000- 0- AP, Proteintech), CD81 mouse monoclonal (66866- 1- Ig, Proteintech), and IBV NP rabbit polyclonal (GTX128538, GeneTex).

Techniques: Virus, Produced, Plasmid Preparation, Purification, Western Blot, Infection, Mutagenesis, Amplification, Residue, RNA Extraction, Clone Assay

Journal: Disease Markers

Article Title: ANP32 Family as Diagnostic, Prognostic, and Therapeutic Biomarker Related to Immune Infiltrates in Hepatocellular Carcinoma

doi: 10.1155/2022/5791471

Figure Lengend Snippet: Expression and diagnostic value of ANP32 family members in hepatocellular carcinoma (HCC). (a) Differential expression of ANP32 family members between HCC tissues and normal liver tissues in the TCGA + GETx cohort. (b) Differential expression of ANP32 family members between HCC tissues and normal liver tissues in paired samples from the TCGA cohort. (c) Expression of ANP32 family members in normal liver tissues, primary HCC tissues, and metastatic HCC tissues by TNM plot. (d) Expression of ANP32 family members in HCC tissues and normal liver tissues by immunohistochemistry. (e) Expression of ANP32 family members in HCC tissues and normal liver tissues by western blot. (f) Quantification of western blot data. (g)–(i) Diagnostic ROC curves of ANP32A (g), ANP32B (h), and ANP32E (i) in the TCGA + GETx cohort.

Article Snippet: The sections were then blocked and stained with antibodies against ANP32A , ANP32B , and ANP32E (dilution 1 : 100), followed by the corresponding secondary antibody and a Streptavidin Biotin Complex kit (Boster BioEngineering, Wuhan, China).

Techniques: Expressing, Diagnostic Assay, Quantitative Proteomics, Immunohistochemistry, Western Blot

Journal: Disease Markers

Article Title: ANP32 Family as Diagnostic, Prognostic, and Therapeutic Biomarker Related to Immune Infiltrates in Hepatocellular Carcinoma

doi: 10.1155/2022/5791471

Figure Lengend Snippet: The association between ANP32A expression and clinical features of HCC patitents in TCGA cohort.

Article Snippet: The sections were then blocked and stained with antibodies against ANP32A , ANP32B , and ANP32E (dilution 1 : 100), followed by the corresponding secondary antibody and a Streptavidin Biotin Complex kit (Boster BioEngineering, Wuhan, China).

Techniques: Expressing

Journal: Disease Markers

Article Title: ANP32 Family as Diagnostic, Prognostic, and Therapeutic Biomarker Related to Immune Infiltrates in Hepatocellular Carcinoma

doi: 10.1155/2022/5791471

Figure Lengend Snippet: ANP32 family member expression was related to Ki-67 . Relationships between Ki-67 and ANP32A (a), ANP32B (b), and ANP32E (c).

Article Snippet: The sections were then blocked and stained with antibodies against ANP32A , ANP32B , and ANP32E (dilution 1 : 100), followed by the corresponding secondary antibody and a Streptavidin Biotin Complex kit (Boster BioEngineering, Wuhan, China).

Techniques: Expressing

Journal: Disease Markers

Article Title: ANP32 Family as Diagnostic, Prognostic, and Therapeutic Biomarker Related to Immune Infiltrates in Hepatocellular Carcinoma

doi: 10.1155/2022/5791471

Figure Lengend Snippet: Univariate and multivariate Cox regression analyses of selected variables on OS.

Article Snippet: The sections were then blocked and stained with antibodies against ANP32A , ANP32B , and ANP32E (dilution 1 : 100), followed by the corresponding secondary antibody and a Streptavidin Biotin Complex kit (Boster BioEngineering, Wuhan, China).

Techniques:

Journal: Disease Markers

Article Title: ANP32 Family as Diagnostic, Prognostic, and Therapeutic Biomarker Related to Immune Infiltrates in Hepatocellular Carcinoma

doi: 10.1155/2022/5791471

Figure Lengend Snippet: Univariate and multivariate Cox regression analyses of selected variables on DSS.

Article Snippet: The sections were then blocked and stained with antibodies against ANP32A , ANP32B , and ANP32E (dilution 1 : 100), followed by the corresponding secondary antibody and a Streptavidin Biotin Complex kit (Boster BioEngineering, Wuhan, China).

Techniques:

Journal: Disease Markers

Article Title: ANP32 Family as Diagnostic, Prognostic, and Therapeutic Biomarker Related to Immune Infiltrates in Hepatocellular Carcinoma

doi: 10.1155/2022/5791471

Figure Lengend Snippet: Gene set enrichment analysis (GSEA) of the ANP32 family. (a) and (b) GSEA for ANP32A based on Reactome pathways and GO. (c) and (d) GSEA for ANP32B based on Reactome pathways and GO. (e) and (f) GSEA for ANP32E based on Reactome pathways and GO.

Article Snippet: The sections were then blocked and stained with antibodies against ANP32A , ANP32B , and ANP32E (dilution 1 : 100), followed by the corresponding secondary antibody and a Streptavidin Biotin Complex kit (Boster BioEngineering, Wuhan, China).

Techniques:

Journal: Disease Markers

Article Title: ANP32 Family as Diagnostic, Prognostic, and Therapeutic Biomarker Related to Immune Infiltrates in Hepatocellular Carcinoma

doi: 10.1155/2022/5791471

Figure Lengend Snippet: Relationships between ANP32 family members and immune characteristics. (a)–(c) Relationship between ANP32 family member expression and immune cell infiltration by ssGSEA. (d)–(f) Relationship between ANP32 family member expression and immune status by ssGSEA. (g) Associations between ANP32A , ANP32B , and ANP32E with immune subtypes in HCC by TISIDB.

Article Snippet: The sections were then blocked and stained with antibodies against ANP32A , ANP32B , and ANP32E (dilution 1 : 100), followed by the corresponding secondary antibody and a Streptavidin Biotin Complex kit (Boster BioEngineering, Wuhan, China).

Techniques: Expressing