Journal: Molecular medicine reports

Article Title: Preparation of a monoclonal antibody against the carcinoembryonic antigen, glypican‑3.

doi: 10.3892/mmr.2019.10019

Figure Lengend Snippet: Figure 3. Expression, purification and identification of recombinant GPC3 protein. (A) Induction expression of recombinant GPC3 protein by SDS‑PAGE analysis. Lane M, protein molecular weight marker; Lane 1, non‑induction of GPC3; lane 2, induction of GPC3. (B) SDS‑PAGE analysis of purification of recombinant GPC3 protein. M, standard protein molecular weight; lane 1, purified protein. (C) Confirmation of the recombinant protein by western blot analysis. The primary antibody was rabbit anti‑His‑tag monoclonal antibody. M, standard protein molecular weight; lane 1, purified protein. GPC3, glypican‑3.

Article Snippet: The recombinant plasmid was transfected into E. coli BL21 (DE3, Novagen; Merck KGaA) with the soluble fusion protein GPC3 expressed following induction with 0.1 mM isopropyl-β-Dthiogalactopyranoside (IPTG; Takara Bio, Inc.), and non-IPTG cells were used as control.

Techniques: Expressing, Purification, Recombinant, Molecular Weight, Marker, Western Blot

Journal: Translational Lung Cancer Research

Article Title: Senescence marker protein 30 inhibits tumor growth by reducing HDAC4 expression in non-small cell lung cancer

doi: 10.21037/tlcr-21-982

Figure Lengend Snippet: Correlation of low SMP30 level with worse overall survival in NSCLC patients. (A) Representative immunohistochemical (IHC) images for SMP30 expression in NSCLC (lung squamous cell carcinoma and lung adenocarcinoma) and adjacent noncancerous tissues. Scale bar, 200 and 20 µm (inset) respectively. (B) Statistical analysis of SMP30 expression in tumor tissue and adjacent noncancerous tissue of 341 NSCLC patients through IHC staining. (C-F) Kaplan-Meier survival analyses of high/low SMP30 expression based on tissue microarray IHC results for 341 NSCLC patients (C), 1,926 NSCLC patients in the Kaplan-Meier plotter database (D), 1,014 NSCLC patients from The Human Protein Atlas (original RNA-seq data from TCGA) (E), 152 NSCLC patients in the early clinical stages, and 189 NSCLC patients in the late clinical stages (F). (G) Statistical analysis of SMP30 expression in different clinical stages and T classifications of 341 NSCLC patients through IHC staining. SMP30, senescence marker protein 30; NSCLC, non-small cell lung cancer; TCGA, The Cancer Genome Atlas.

Article Snippet: Using paraffin-embedded tissue microarray containing 341 pairs NSCLC and matched adjacent normal lung tissues, IHC staining was conducted using the primary antibodies of anti- SMP30 (1:50, 17947-1-AP, Proteintech), and standard protocols were followed as previously described ( 30 ).

Techniques: Immunohistochemical staining, Expressing, Immunohistochemistry, Microarray, RNA Sequencing, Marker

Journal: Translational Lung Cancer Research

Article Title: Senescence marker protein 30 inhibits tumor growth by reducing HDAC4 expression in non-small cell lung cancer

doi: 10.21037/tlcr-21-982

Figure Lengend Snippet: Association of SMP30 expression with clinicopathologic parameters of patients with NSCLC

Article Snippet: Using paraffin-embedded tissue microarray containing 341 pairs NSCLC and matched adjacent normal lung tissues, IHC staining was conducted using the primary antibodies of anti- SMP30 (1:50, 17947-1-AP, Proteintech), and standard protocols were followed as previously described ( 30 ).

Techniques: Expressing

Journal: Translational Lung Cancer Research

Article Title: Senescence marker protein 30 inhibits tumor growth by reducing HDAC4 expression in non-small cell lung cancer

doi: 10.21037/tlcr-21-982

Figure Lengend Snippet: Univariate and multivariate analyses of the correlation between clinicopathological variables for survival of patients with non-small cell lung cancer

Article Snippet: Using paraffin-embedded tissue microarray containing 341 pairs NSCLC and matched adjacent normal lung tissues, IHC staining was conducted using the primary antibodies of anti- SMP30 (1:50, 17947-1-AP, Proteintech), and standard protocols were followed as previously described ( 30 ).

Techniques: Expressing

Journal: Translational Lung Cancer Research

Article Title: Senescence marker protein 30 inhibits tumor growth by reducing HDAC4 expression in non-small cell lung cancer

doi: 10.21037/tlcr-21-982

Figure Lengend Snippet: Correlation of low SMP30 level with worse overall survival in both LUAD and LUSC patients. (A,E) Kaplan-Meier survival analyses of high/low SMP30 expression in 719 LUAD patients and 524 LUSC patients, respectively, based on the Kaplan-Meier plotter database. (B-D,F-H) Kaplan-Meier survival analyses of high/low SMP30 expression in 131 LUAD patients and 210 LUSC patients based on the tissue microarray immunohistochemical results. SMP30, senescence marker protein 30; HR, hazard ratio; LUAD, lung adenocarcinoma; LUSC, lung squamous cell carcinoma.

Article Snippet: Using paraffin-embedded tissue microarray containing 341 pairs NSCLC and matched adjacent normal lung tissues, IHC staining was conducted using the primary antibodies of anti- SMP30 (1:50, 17947-1-AP, Proteintech), and standard protocols were followed as previously described ( 30 ).

Techniques: Expressing, Microarray, Immunohistochemical staining, Marker

Journal: Translational Lung Cancer Research

Article Title: Senescence marker protein 30 inhibits tumor growth by reducing HDAC4 expression in non-small cell lung cancer

doi: 10.21037/tlcr-21-982

Figure Lengend Snippet: Effect of SMP30 overexpression on proliferative ability of A549 and H1299 cells, and tumor growth in H1299 cell tumor xenografts. (A) Relative protein levels of SMP30 in cell lines as determined by Western blot. β-actin was used as a loading control. (B) Representative Western blot results of Flag, SMP30, c-Myc and CyclinD1 in LV-SMP30 A549 and LV-SMP30 H1299 cells. (C) Growth curves of the A549 and H1299 cells. Cell viability was detected by CCK-8 assay and expressed as optical density values. (D) Representative images and results of the colony formation assay. Colonies were visualized by crystal violet staining. (E) Representative images (×20) and statistical analysis of EdU incorporation assay. The results are presented as the ratio between the number of EdU-positive cells (red fluorescence) and the total number of Hoechst 33342-stained cells (blue fluorescence). (F) Representative results of tumor weight changes after subcutaneous injection of H1299 cells. Photographs show tumor xenograft morphologies in the LV-control and LV-SMP30 H1299 groups. (G) Representative Western blot results of Flag, SMP30, c-Myc and CyclinD1 in H1299 xenograft tumor tissues. β-actin was used as a Western blot loading control. The values below the Western blot band represent the relative gray values. All the data are shown as the mean ± SD. Student’s t-test, *, P<0.05 vs. LV-control group. LV, lentivirus; SMP30, senescence marker protein 30.

Article Snippet: Using paraffin-embedded tissue microarray containing 341 pairs NSCLC and matched adjacent normal lung tissues, IHC staining was conducted using the primary antibodies of anti- SMP30 (1:50, 17947-1-AP, Proteintech), and standard protocols were followed as previously described ( 30 ).

Techniques: Over Expression, Western Blot, Control, CCK-8 Assay, Colony Assay, Staining, Fluorescence, Injection, Marker

Journal: Translational Lung Cancer Research

Article Title: Senescence marker protein 30 inhibits tumor growth by reducing HDAC4 expression in non-small cell lung cancer

doi: 10.21037/tlcr-21-982

Figure Lengend Snippet: Effect of HDAC4 level of SMP30 overexpression both in vivo and in vitro, and effect of HDAC4 overexpression on NSCLC cell proliferation. (A) Representative Western blot results of HDAC4, HDAC5, HDAC7 and HDAC9 in LV-SMP30 A549 and LV-SMP30 H1299 cells. (B) Representative Western blot results of HDAC4, HDAC5, HDAC7 and HDAC9 in H1299 xenograft tumor tissues. (C) Relative protein levels of HDAC4 in cell lines as determined by Western blot. (D) Representative Western blot results of Flag, HDAC4, c-Myc and CyclinD1 in LV-HDAC4 A549 and LV-HDAC4 H1299 cells. (E) Representative images and results of colony formation assay. Colonies were visualized by crystal violet staining. (F) Representative images (×20) and statistical analysis of EdU incorporation assay. The results are presented as the ratio between the number of EdU-positive cells (red fluorescence) and the total number of Hoechst 33342-stained cells (blue fluorescence). β-actin was used as a Western blot loading control. The values below the Western blot band represent the relative gray values. All the data are shown as the mean ± SD. Student’s t-test, *, P<0.05 vs. LV-control group. HDAC histone deacetylase; LV, lentivirus; SMP30, senescence marker protein 30; SD, standard deviation; NSCLC, non-small cell lung cancer.

Article Snippet: Using paraffin-embedded tissue microarray containing 341 pairs NSCLC and matched adjacent normal lung tissues, IHC staining was conducted using the primary antibodies of anti- SMP30 (1:50, 17947-1-AP, Proteintech), and standard protocols were followed as previously described ( 30 ).

Techniques: Over Expression, In Vivo, In Vitro, Western Blot, Colony Assay, Staining, Fluorescence, Control, Histone Deacetylase Assay, Marker, Standard Deviation

Journal: Translational Lung Cancer Research

Article Title: Senescence marker protein 30 inhibits tumor growth by reducing HDAC4 expression in non-small cell lung cancer

doi: 10.21037/tlcr-21-982

Figure Lengend Snippet: Essential requirement for HDAC4 in SMP30-mediated NSCLC cell proliferation. (A) Representative Western blot results of Flag, SMP30, HDAC4 and c-Myc and CyclinD1. (B) Representative images and results of colony formation assay. Colonies were visualized by crystal violet staining. (C) Representative images (×20) and statistical analysis of EdU incorporation assay. The results are presented as the ratio between the number of EdU-positive cells (red fluorescence) and the total number of Hoechst 33342-stained cells (blue fluorescence). (D) Representative results for tumor weight changes after subcutaneous injection of H1299 cells. Photographs show tumor xenograft morphologies in the LV-SMP30 + LV-HDAC4 and LV-SMP30 + LV-control H1299 groups. (E) Representative Western blot results of SMP30, HDAC4, c-Myc and CyclinD1 in H1299 xenograft tumor tissues. β-actin was used as a Western blot loading control. The values below the Western blot band represent the relative gray values. All the data are shown as the mean ± SD. Student’s t-test, *, P<0.05 vs. the LV-SMP30 control group; #, P<0.05 vs. the LV-SMP30 group; &, P<0.05 vs. the LV-SMP30 control + LV-HDAC4 group. HDAC, histone deacetylase; LV, lentivirus; SMP30, senescence marker protein 30; SD, standard deviation; NSCLC, non-small cell lung cancer.

Article Snippet: Using paraffin-embedded tissue microarray containing 341 pairs NSCLC and matched adjacent normal lung tissues, IHC staining was conducted using the primary antibodies of anti- SMP30 (1:50, 17947-1-AP, Proteintech), and standard protocols were followed as previously described ( 30 ).

Techniques: Western Blot, Colony Assay, Staining, Fluorescence, Injection, Control, Histone Deacetylase Assay, Marker, Standard Deviation

Journal: Translational Lung Cancer Research

Article Title: Senescence marker protein 30 inhibits tumor growth by reducing HDAC4 expression in non-small cell lung cancer

doi: 10.21037/tlcr-21-982

Figure Lengend Snippet: Effect of pharmacologic inhibition of HDAC4 on proliferative abilities of H1299 cells. LV-control and LV-HDAC4 H1299 cells were pretreated with SAHA or TMP269 for 48 h. (A,B) Representative Western blot results of HDAC4, c-Myc, and CyclinD1. (C,D) Representative images and results of colony formation assay. Colonies were visualized by crystal violet staining. (E,F) Representative images (×20) and statistical analysis of EdU incorporation assay. The results are presented as the ratio between the number of EdU-positive cells (red fluorescence) and the total number of Hoechst 33342-stained cells (blue fluorescence). *, P<0.05 vs. the LV-control +SAHA/TMP269 0 µM group; #, P<0.05 vs. the LV-control + SAHA1.25 µM/TMP269 40 µM group; &, P<0.05 vs. the LV-HDAC4 + SAHA/TMP269 0 µM group. HDAC, histone deacetylase; LV, lentivirus; SMP30, senescence marker protein 30.

Article Snippet: Using paraffin-embedded tissue microarray containing 341 pairs NSCLC and matched adjacent normal lung tissues, IHC staining was conducted using the primary antibodies of anti- SMP30 (1:50, 17947-1-AP, Proteintech), and standard protocols were followed as previously described ( 30 ).

Techniques: Inhibition, Control, Western Blot, Colony Assay, Staining, Fluorescence, Histone Deacetylase Assay, Marker