Journal: Stem Cell Research & Therapy

Article Title: Senescent lung-resident mesenchymal stem cells drive pulmonary fibrogenesis through FGF-4/FOXM1 axis

doi: 10.1186/s13287-024-03866-2

Figure Lengend Snippet: FOXM1 plays an essential role in FGF-4-induced pulmonary fibroblasts activation and pulmonary fibrogenesis. ( A ) Representative images of coimmunostaining of FOXM1 and α-SMA in lung tissues from patients with IPF. ( B ) Representative images of coimmunostaining of FOXM1 and α-SMA in lung tissues from bleomycin (BLM)-treated mice. ( C ) Western blot analysis of FOXM1 expression in pulmonary fibroblasts isolated from BLM-treated mice. ( D ) Western blot analysis of protein levels of COL1A1, FOXM1 and α-SMA in FGF-4-treated pulmonary fibroblasts followed with the treatment of Thiostrepton (Thi, 10 µM). ( E ) Co-IP analysis of β-catenin-FOXM1 interaction in pulmonary fibroblasts treated as in D. ( F , G ) Immunofluorescence analysis the expression of FOXM1 ( F ), α-SMA and COL1A1 ( G ) in pulmonary fibroblasts treated as in D. ( H ) The migration ability assay of pulmonary fibroblasts treated as in D. n = 3, ** P < 0.01. ( I ) Haematoxylin and eosin ( H & E ) staining, Masson’s trichrome staining of representative lung sections ( n = 6) from BLM-treated mice injected with or without Thi (30 mg/kg). ( J ) Western blot analysis of α-SMA, β-catenin and FOXM1 expression in the lung tissues from mice treated as in I. ( K ) Representative images of coimmunostaining of α-SMA and COL1A1in lung tissues from mice treated as in I. ( L ) Representative images of coimmunostaining of FOXM1 and α-SMA in lung tissues from mice treated as in I

Article Snippet: To explore the role of FOXM1 in pulmonary fibrosis, mice were treated with thiostrepton (HY-B0990, MedChem Express, San Diego, CA), which was known as an inhibitor of FOXM1 [ ].

Techniques: Activation Assay, Western Blot, Expressing, Isolation, Co-Immunoprecipitation Assay, Immunofluorescence, Migration, Staining, Injection

Journal: Acta pharmaceutica Sinica. B

Article Title: Protocatechuic aldehyde protects cardiomycoytes against ischemic injury via regulation of nuclear pyruvate kinase M2.

doi: 10.1016/j.apsb.2021.03.021

Figure Lengend Snippet: Figure 2 Identification of PKM2 as a direct binding target for PCA. (A) The employed drug affinity responsive target stabilization (DARTS) assay detects a marked increase in w60 kD band upon PCA incubation in pronase digested H9C2 cell lysates. (B) Immunoblot analysis of PKM2 in pronase-digested cell lysate (n Z 3). (C) Recombinant PKM2 in pronase-digested H9C2 cell lysates. PKM2 degradation in H9C2 cell lysates (D) and in intact cells (E) (n Z 3). (F) Surface plasmon resonance (SPR) binding curve fit to a Kd Z 5.76 nmol/L for PCA and PKM2 (n Z 3). Results are shown as mean SD; *P < 0.05, ***P < 0.001.

Article Snippet: Antibodies against GAPDH (60004-1-Ig), b-catenin (51067-2-AP, 66379-1-Ig), PCNA (10205-2-AP), caspase 3 (66470-2-Ig), BAX (60267-1-Ig), BCL-2 (60178-1-Ig), PKM2 (60268-1-Ig), b-tubulin (10068-1-AP) and TCF4 (22337-1-AP) were obtained from Proteintech (Wuhan, China).

Techniques: Binding Assay, Incubation, Western Blot, Recombinant, SPR Assay

Journal: Acta pharmaceutica Sinica. B

Article Title: Protocatechuic aldehyde protects cardiomycoytes against ischemic injury via regulation of nuclear pyruvate kinase M2.

doi: 10.1016/j.apsb.2021.03.021

Figure Lengend Snippet: Figure 3 PCA PKM2-dependently protects cardiomyocytes. Primary neonatal rat ventricular myocytes (NRVMs) were cultured in glucose-free DMEM under 1% O2 (OGD) for 4 h in the presence of protocatechuic aldehyde (PCA). (A) Cell survival in NRVMs exposed to OGD for 6 h (n Z 6). (B) Intracellular ROS production (n Z 6). (C) Representative images of mitochondrial permeability transition pore (mPTP) and quantification of relative fluorescence intensity (n Z 6, one of three independent experiments). Scale bar: 10 mm. (D) Mitochondrial fission was detected by Mito-Tracker Red (one of 3 independent experiments) and quantification analysis. Scale bar: 10 mm. (E) Caspase 3 activity in NRVMs (n Z 6). (F) Representative Western blots of BAX, BCL-2, caspase 3, cleaved caspase 3 (c-caspase 3) and the ratio of BAX/BCL-2 (n Z 3). (G) Representative images of TUNEL staining and quantification analysis the percentage of apoptosis cells. Scale bars, 100 mm (n Z 3). (H) Data of Annexin V/PI double staining flow cytometry; the percentage for each panel indicates the percentage of apoptotic cells and quantification of the percentage of apoptotic cells (n Z 3). Results are shown as mean SD; *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Antibodies against GAPDH (60004-1-Ig), b-catenin (51067-2-AP, 66379-1-Ig), PCNA (10205-2-AP), caspase 3 (66470-2-Ig), BAX (60267-1-Ig), BCL-2 (60178-1-Ig), PKM2 (60268-1-Ig), b-tubulin (10068-1-AP) and TCF4 (22337-1-AP) were obtained from Proteintech (Wuhan, China).

Techniques: Cell Culture, Permeability, Activity Assay, Western Blot, TUNEL Assay, Staining, Double Staining, Cytometry

Journal: Acta pharmaceutica Sinica. B

Article Title: Protocatechuic aldehyde protects cardiomycoytes against ischemic injury via regulation of nuclear pyruvate kinase M2.

doi: 10.1016/j.apsb.2021.03.021

Figure Lengend Snippet: Figure 4 PCA promotes the nuclear localization of PKM2. Primary neonatal rat ventricular myocytes (NRVMs) were exposed to 1% O2 in glucose-free DMEM (OGD) for 4 h in the presence of protocatechuic aldehyde (PCA). (A) PKM2 activity in NRVMs (n Z 6). (B) Immuno- fluorescence image of endogenous PKM2 (one of three independent experiments. Scale bar: 10 mm. (C) Nuclear PKM2 protein expression (n Z 3). (D) Cytoplasmic PKM2 protein expression (n Z 3). (E) Total PKM2 protein expression (n Z 3). (F) Docking analysis illustrates the interaction between PCA and PKM2. The residues that are likely to participate in the interactions with PCA are labeled. (G) Representative co- immunoprecipitation (co-IP) analysis of PKM2 and SIRT6 in the NRVMs (n Z 3). (H) Representative co-IP of PKM2 and b-catenin in the NRVMs (n Z 3). Results are shown as mean SD; *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Antibodies against GAPDH (60004-1-Ig), b-catenin (51067-2-AP, 66379-1-Ig), PCNA (10205-2-AP), caspase 3 (66470-2-Ig), BAX (60267-1-Ig), BCL-2 (60178-1-Ig), PKM2 (60268-1-Ig), b-tubulin (10068-1-AP) and TCF4 (22337-1-AP) were obtained from Proteintech (Wuhan, China).

Techniques: Activity Assay, Expressing, Labeling, Immunoprecipitation, Co-Immunoprecipitation Assay

Journal: Acta pharmaceutica Sinica. B

Article Title: Protocatechuic aldehyde protects cardiomycoytes against ischemic injury via regulation of nuclear pyruvate kinase M2.

doi: 10.1016/j.apsb.2021.03.021

Figure Lengend Snippet: Figure 6 The myocardial protection effect of PCA is dependent on PKM2. Mice were orally administrated with protocatechuic aldehyde (PCA) for 3 weeks after coronary artery ligation. (A) Representative photomicrographs for HE staining of cardiac tissue sections (n Z 6), Scale bar, 50 mm. (B) Representative photomicrographs for masson staining of cardiac tissue sections (n Z 6). Scale bar, 50 mm. (C) 4-HNE contents in the heart (n Z 6). (D) Representative M mode images of echocardiography from the assayed groups under treatments as indicated (n Z 6). (E) Ejection fractions (EF) and shortening fraction (FS) in mice (n Z 6). (F) Representative co-IP analysis of PKM2 and b-catenin in the heart (n Z 3). The mRNA levels of Myc (G), Ccnd1 (H) and Sgk1 (I) in the heart (n Z 6). (J) Immunocytochemical staining of BAX, BCL-2 and c- caspase 3 protein expression (n Z 6), scale bar Z 50 mm. (K) Representative TUNEL staining images and quantification of TUNEL positive cells (n Z 6). Results are shown as mean SD; *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Antibodies against GAPDH (60004-1-Ig), b-catenin (51067-2-AP, 66379-1-Ig), PCNA (10205-2-AP), caspase 3 (66470-2-Ig), BAX (60267-1-Ig), BCL-2 (60178-1-Ig), PKM2 (60268-1-Ig), b-tubulin (10068-1-AP) and TCF4 (22337-1-AP) were obtained from Proteintech (Wuhan, China).

Techniques: Ligation, Staining, Co-Immunoprecipitation Assay, Expressing, TUNEL Assay

Journal:

Article Title: Formation of Wild-Type and Chimeric Influenza Virus-Like Particles following Simultaneous Expression of Only Four Structural Proteins

doi: 10.1128/JVI.75.13.6154-6165.2001

Figure Lengend Snippet: (A) Diagram of quadruple baculovirus transfer vector encoding influenza virus genes HA, NA, M1, and M2. This vector was transfected together with linearized baculovirus DNA into Sf9 insect cells to generate the quadruple recombinant (HA/Q28). Replacement of the HA with a gene encoding a full-length sequence of the VSV-G protein or chimera VSV-G/HA (see below) generated VSV-G/Q or VSV-G/HA/Q, respectively. (B) Structure of the VSV-G/HA chimera. The 11 aa of the cytoplasmic tail, 26 aa of the transmembrane domain, and 5 aa of the ectodomain of the influenza virus HA were fused in frame with the ectodomain of the VSV-G surface glycoprotein. This hybrid gene was used to replace the HA from the quadruple transfer vector (panel A) and generate the VSV-G/HA/Q gene.

Article Snippet: The antibodies used in this work were obtained from the following sources: rat monoclonal anti-HA (clone 3F10), mouse monoclonal anti-HA (clone 12CA5), and mouse monoclonal anti-VSV-G (clone P5D4) were obtained from Roche Molecular Biochemicals (Indianapolis, Ind.); mouse monoclonal anti-M1 (clone GA2B) was from Serotec (Raleigh, N.C.); goat polyclonal anti-M1 was from Accurate Chemical & Scientific Corp. (Westbury, N.Y.); mouse monoclonal anti-NA (clone ST9D2) was from Argene Inc. (Massapequa, N.Y.); mouse monoclonal anti-M2 was from Mt.

Techniques: Plasmid Preparation, Transfection, Recombinant, Sequencing, Generated

Journal:

Article Title: Formation of Wild-Type and Chimeric Influenza Virus-Like Particles following Simultaneous Expression of Only Four Structural Proteins

doi: 10.1128/JVI.75.13.6154-6165.2001

Figure Lengend Snippet: Western blot analysis of the influenza virus and VSV-G proteins expressed in Sf9 cells infected by quadruple baculovirus recombinants (HA/Q28 or VSV-G/Q). (A) Expression of the influenza virus proteins HA, NA, and M2 in the cell pellet (lane 2) and culture supernatant (lane 3) was evaluated with a mixture of anti-HA, anti-M1, and anti-M2 monoclonal antibodies. Uninfected Sf9 cells and influenza virus A/Udorn-infected MDCK cells were used as controls (lanes 1 and 4, respectively). (B) Expression of VSV-G as well as influenza virus M1 and M2 proteins in Sf9 cells infected with VSV-G/Q (full-length G) was evaluated in cell pellets (lane 2) and culture supernatants (lane 3) by using a mixture of anti-G, anti-M1, and anti-M2 monoclonal antibodies. Uninfected Sf9 cells (lane 1), and VSV-infected BHK cells (lane 4) and influenza virus A/Udorn-infected MDCK cells (lane 5) were used as negative and positive controls, respectively.

Article Snippet: The antibodies used in this work were obtained from the following sources: rat monoclonal anti-HA (clone 3F10), mouse monoclonal anti-HA (clone 12CA5), and mouse monoclonal anti-VSV-G (clone P5D4) were obtained from Roche Molecular Biochemicals (Indianapolis, Ind.); mouse monoclonal anti-M1 (clone GA2B) was from Serotec (Raleigh, N.C.); goat polyclonal anti-M1 was from Accurate Chemical & Scientific Corp. (Westbury, N.Y.); mouse monoclonal anti-NA (clone ST9D2) was from Argene Inc. (Massapequa, N.Y.); mouse monoclonal anti-M2 was from Mt.

Techniques: Western Blot, Infection, Expressing

Journal:

Article Title: Formation of Wild-Type and Chimeric Influenza Virus-Like Particles following Simultaneous Expression of Only Four Structural Proteins

doi: 10.1128/JVI.75.13.6154-6165.2001

Figure Lengend Snippet: Immunofluorescence analysis of Sf9 cells infected with a quadruple baculovirus recombinant (HA/Q28). Expression of the influenza virus proteins HA, M1, or HA/M1 (A) and HA, NA, or HA/NA (B) was detected with a combination of anti-HA and anti-M1 or anti-HA and anti-NA antibodies as described in Materials and Methods. (C) M2 expression was examined with a rabbit anti-M2 peptide antibody. (D) Sf9 cells infected with VSV/Q chimeric baculovirus recombinant were used to evaluate expression of the VSV-G protein.

Article Snippet: The antibodies used in this work were obtained from the following sources: rat monoclonal anti-HA (clone 3F10), mouse monoclonal anti-HA (clone 12CA5), and mouse monoclonal anti-VSV-G (clone P5D4) were obtained from Roche Molecular Biochemicals (Indianapolis, Ind.); mouse monoclonal anti-M1 (clone GA2B) was from Serotec (Raleigh, N.C.); goat polyclonal anti-M1 was from Accurate Chemical & Scientific Corp. (Westbury, N.Y.); mouse monoclonal anti-NA (clone ST9D2) was from Argene Inc. (Massapequa, N.Y.); mouse monoclonal anti-M2 was from Mt.

Techniques: Immunofluorescence, Infection, Recombinant, Expressing

Journal:

Article Title: Formation of Wild-Type and Chimeric Influenza Virus-Like Particles following Simultaneous Expression of Only Four Structural Proteins

doi: 10.1128/JVI.75.13.6154-6165.2001

Figure Lengend Snippet: Analysis of VLP formation by iodixanol gradient centrifugation. Concentrated supernatants of HA/Q28- or VSV-G/Q recombinant-infected Sf9 cells were layered on top of an iodixanol density gradient and spun at 200,000 × g for 3.5 h. Fractions were collected from the bottom of the tube, resolved by SDS-PAGE, and transferred onto nitrocellulose. (A) Fractions from HA/Q28 were probed with a mixture of anti-HA and anti-M1 monoclonal antibodies. Lanes 1 to 8, fractions collected from top to bottom of the tube; lane 9, influenza virus A/Udorn-infected MDCK cells as control. (B) Fractions from VSV-G/Q were probed with a mixture of anti-VSV-G, anti-M1, and anti-M2 monoclonal antibodies. Lanes 1 to 8, fractions collected from the top to the bottom of the tube; lane 9, VSV-infected BHK cells and influenza virus-infected MDCK cells combined as control. (C) Concentrated supernatants of Sf9 cells infected with the M1 single baculovirus recombinant were purified as above and probed with anti-M1 antibody. Lanes 1 to 8, gradient fractions collected from the top to the bottom of the tube; lane 9, influenza virus-infected MDCK cells (control).

Article Snippet: The antibodies used in this work were obtained from the following sources: rat monoclonal anti-HA (clone 3F10), mouse monoclonal anti-HA (clone 12CA5), and mouse monoclonal anti-VSV-G (clone P5D4) were obtained from Roche Molecular Biochemicals (Indianapolis, Ind.); mouse monoclonal anti-M1 (clone GA2B) was from Serotec (Raleigh, N.C.); goat polyclonal anti-M1 was from Accurate Chemical & Scientific Corp. (Westbury, N.Y.); mouse monoclonal anti-NA (clone ST9D2) was from Argene Inc. (Massapequa, N.Y.); mouse monoclonal anti-M2 was from Mt.

Techniques: Gradient Centrifugation, Recombinant, Infection, SDS Page, Purification

Journal:

Article Title: Formation of Wild-Type and Chimeric Influenza Virus-Like Particles following Simultaneous Expression of Only Four Structural Proteins

doi: 10.1128/JVI.75.13.6154-6165.2001

Figure Lengend Snippet: (A) Comparative Western blot of M1 protein released from influenza virus A/Udorn-infected MDCK cells or from Sf9 cells infected with the quadruple recombinant HA/Q28 or the M1 single recombinant. MDCK cells (5 × 106) were infected with influenza virus A/Udorn (MOI, 1) for 24 h. Equivalent numbers of Sf9 cells were infected with either HA/Q28 or M1 single recombinant (MOI, 5) for 72 h. Supernatants were concentrated and particles were purified by iodixanol gradient centrifugation. Fractions 2 and 3 were pooled, and M1 protein was immunoprecipitated with anti-M1 monoclonal antibody twice to ensure complete recovery. Samples were resolved by SDS-PAGE, transferred to nitrocellulose, and probed with anti-M1 polyclonal antibody. Different amounts of each sample were analyzed to determine the linear range of the detecting color reaction. Lanes 1, 2, and 3 show the relative amount of M1 protein immunoprecipitated and detected by Western blotting from purified influenza virions, HA/Q 28, and M1 particles, respectively. We used NIH Image 1.54 to compare the relative amount of M1 protein detected in each sample. (B) Relative amounts of M1 and VSV-G proteins detected in culture supernatants of Sf9 cells infected with VSV-G/Q or VSV-G/HA/Q. Equivalent numbers of Sf9 cells (5 × 106) were infected with either VSV-G/Q or VSV-G/HA/Q quadruple recombinants (MOI, 5). Culture supernatants were concentrated, and the relative amounts of M1 and VSV-G or VSV-G/HA chimera were measured as described above. Lanes 1 and 2 show the M1 and VSV-G or VSV-G/HA chimeric proteins released from Sf9 cells infected with VSV-G/Q or VSV-G/HA/Q quadruple recombinants, respectively.

Article Snippet: The antibodies used in this work were obtained from the following sources: rat monoclonal anti-HA (clone 3F10), mouse monoclonal anti-HA (clone 12CA5), and mouse monoclonal anti-VSV-G (clone P5D4) were obtained from Roche Molecular Biochemicals (Indianapolis, Ind.); mouse monoclonal anti-M1 (clone GA2B) was from Serotec (Raleigh, N.C.); goat polyclonal anti-M1 was from Accurate Chemical & Scientific Corp. (Westbury, N.Y.); mouse monoclonal anti-NA (clone ST9D2) was from Argene Inc. (Massapequa, N.Y.); mouse monoclonal anti-M2 was from Mt.

Techniques: Western Blot, Infection, Recombinant, Purification, Gradient Centrifugation, Immunoprecipitation, SDS Page

Journal: iScience

Article Title: Suv4-20h2 protects against influenza virus infection by suppression of chromatin loop formation

doi: 10.1016/j.isci.2021.102660

Figure Lengend Snippet: Deletion of Suv4-20h2 enhances influenza viral replication (A) WT mice were intratracheally infected with influenza virus (PR8 virus, 100 FFU), and lung tissues were sampled at the indicated time points. Histone was extracted from the lung tissues and was subjected to H4-tail proteomics analysis. Relative expression levels of H4K20me1, H4K20me2, and H4K20me3 to H4 were shown (n = 5 per group). (B–D) MEFs obtained from WT, Suv4-20h1 KO (h1 KO), Suv4-20h2 KO (h2 KO) and Suv4-20h1/Suv4-20h2 double KO (dKO) mice were infected with mock (Flu-) or influenza virus (MOI: 0.5) (Flu+). Cells were fixed and stained with either H4K20me1, H4K20me2 or H4K20me3 Abs, and Hoechst. Data are from three separate experiments. Representative staining is shown. Scale bars: 10 μm (B). Culture supernatant and cells were sampled at 8 hr after infection. Virus titer assessed by FFU in supernatant is shown (C, upper panel). NP mRNA expression of the cells is shown (C, lower panel). Data are from three separate experiments. ∗p < 0.05 compared to WT. Infected cells were fixed, and virus NP protein was stained (green). Nuclei were counterstained with Hoechst (blue). Representative staining is shown. Scale bars: 50 μm (D, left panel). The percentages of NP-positive in total of Hoechst positive are shown (D, right panel). ∗∗p < 0.01 compared to WT. (E) WT and dKO MEFs were infected with influenza virus (MOI: 0.5) and sampled at the indicated time point. Western blots of virus PB2, NP and M1 proteins, along with GAPDH are shown. Data were reproduced in three experiments. The data in (A, C and D) (right panel) are presented as means ± s.e.m.. Statistical analysis were carried out using analysis of variance with Bonferroni post t tests.

Article Snippet: Human SET domain of SUV4-20h2 (2-281) (Active motif) was pre-incubated with virus NP or M1 protein (SinoBiological) (0, 0.05, 0.5, or 1.0 μg) at room temperature for 1 hr and then incubated with polynucleosomes (Active motif) (1.0 μg) for additional 3 hr.

Techniques: Infection, Expressing, Staining, Western Blot

Journal: iScience

Article Title: Suv4-20h2 protects against influenza virus infection by suppression of chromatin loop formation

doi: 10.1016/j.isci.2021.102660

Figure Lengend Snippet: Suv4-20h2 interacts with virus and host nuclear proteins (A) dKO cells transduced with EGFP-tagged empty vector (EV), Suv4-20h2 full length (h2-FL), SET domain (h2-SET) or Clamp domain (h2-Clamp; left panel) were infected with influenza virus (MOI: 0.5) for 8 hr h2-EGFP and virus NP protein were co-stained. Nuclei were counterstained with Hoechst. The percentages of NP-positive in total of the h2 EGFP-positive are shown. Data are from three separate experiments. ∗∗P < 0.01 between the groups. (B) WT, dKO, and dKO MEFs overexpressed with Suv4-20h2 were infected with influenza virus (MOI: 0.5). Culture supernatant and cells were sampled at 8 hr after infection. Virus titer of FFU in supernatant is shown (B, left panel). NP mRNA expression of the cells is shown (B, right panel). Data are from three separate experiments. ∗p < 0.05, ∗∗p < 0.01 between the groups. (C) dKO MEFs transduced with AM-tagged Suv4-20h2 were infected with influenza virus (MOI: 0.5) for 8 hr. Nuclear extracts were prepared from the cells, and were immunoprecipitated (IP) with AM Ab, and IP product was subjected to a stable isotope labeling (SILAC) & LC-MS/MS analysis. Virus NP and PB2 proteins were identified in this analysis. (D) Suv4-20h2 Flag knock-in mouse embryonic stem (mES) cells were mock (Flu-) or infected with influenza virus (MOI: 0.5) (Flu+) for 8 hr. Nuclear extracts were prepared from the cells, and were immunoprecipitated with Flag Ab. Western blotting shows that virus NP and PB2 proteins were co-precipitated with Suv4-20h2. Data were reproduced in three experiments. (E) Purified recombinant viral NP and PB2 proteins and GST-fusion proteins to full length (FL), SET, and Clamp domain of Suv4-20h2, were used in a pull-down assay. Virus NP and PB2 proteins interacted with FL and SET domain of Suv4-20h2. Data were reproduced in three experiments. (F) Human SET domain of SUV4-20H2 (2-281) was pre-incubated with virus NP protein (0, 0.05, 0.5, or 1.0 μg) (left panel) or M1 protein (0, 0.05, 0.5, or 1.0 μg) (right panel) at room temperature for 1hr and then incubated with polynucleosomes (1.0 μg) for additional 3hr. Protein samples were subjected to Western blot analysis using specific antibodies. H4K20me3 was suppressed in a dose-dependent manner by NP protein. Data were reproduced in three experiments. The data in (A and B) are presented as means ± s.e.m.. Statistical analysis were carried out using analysis of variance with Bonferroni post t tests.

Article Snippet: Human SET domain of SUV4-20h2 (2-281) (Active motif) was pre-incubated with virus NP or M1 protein (SinoBiological) (0, 0.05, 0.5, or 1.0 μg) at room temperature for 1 hr and then incubated with polynucleosomes (Active motif) (1.0 μg) for additional 3 hr.

Techniques: Transduction, Plasmid Preparation, Infection, Staining, Expressing, Immunoprecipitation, Labeling, Liquid Chromatography with Mass Spectroscopy, Knock-In, Western Blot, Purification, Recombinant, Pull Down Assay, Incubation

Journal: iScience

Article Title: Suv4-20h2 protects against influenza virus infection by suppression of chromatin loop formation

doi: 10.1016/j.isci.2021.102660

Figure Lengend Snippet:

Article Snippet: Human SET domain of SUV4-20h2 (2-281) (Active motif) was pre-incubated with virus NP or M1 protein (SinoBiological) (0, 0.05, 0.5, or 1.0 μg) at room temperature for 1 hr and then incubated with polynucleosomes (Active motif) (1.0 μg) for additional 3 hr.

Techniques: Recombinant, Transfection, Protease Inhibitor, Western Blot, Purification, Clone Assay, Fractionation, Sequencing, Software

Journal: Genetics Research

Article Title: Impact of Extracellular Matrix-Related Genes on the Tumor Microenvironment and Prognostic Indicators in Esophageal Cancer: A Comprehensive Analytical Study

doi: 10.1155/2024/3577395

Figure Lengend Snippet: Identification of the ECM-related genes associated with patients survival. Notes . (a) Univariate Cox regression analysis was utilized to identify the prognosis-related genes with P < 0.05; (b) LASSO regression analysis; (c) multivariate cox regression analysis based on the genes screened by LASSO regression; (d–i) clinical correlation of IBSP, LINGO4, COL26A1, MMP12, KLK4, RTBDN, TENM1, GDF15, and RUNX1.

Article Snippet: Equal amounts of protein were loaded onto SDS-PAGE gels, transferred to PVDF membranes, and probed with antibodies specific to TENM1 (proteintech, 21696-1-AP, 1 : 1500) and GAPDH (proteintech, 60004-1-Ig, 1 : 10000).

Techniques:

Journal: Genetics Research

Article Title: Impact of Extracellular Matrix-Related Genes on the Tumor Microenvironment and Prognostic Indicators in Esophageal Cancer: A Comprehensive Analytical Study

doi: 10.1155/2024/3577395

Figure Lengend Snippet: Expression level of TENM1 in EC cells. Notes . (a) The mRNA level of TENM1 in EC and normal cells; (b) the protein level of TENM1 in EC and normal cells.

Article Snippet: Equal amounts of protein were loaded onto SDS-PAGE gels, transferred to PVDF membranes, and probed with antibodies specific to TENM1 (proteintech, 21696-1-AP, 1 : 1500) and GAPDH (proteintech, 60004-1-Ig, 1 : 10000).

Techniques: Expressing