Journal: British Journal of Cancer

Article Title: Gene expression reveals two distinct groups of anal carcinomas with clinical implications

doi: 10.1038/sj.bjc.6604285

Figure Lengend Snippet: ( A ) Hierarchical clustering of the unsupervised analysis on ( Aa ) the selection of 8449 probes, ( Ab ) the 337 individual genes defined by the T -test, ( Ac ) the 363 individual genes defined by the SAM analysis together with collections of genes described by others relevant to our study and included within the group of 8449 probes, ( Ad ) HPV16-induced gene expression ( Alazawi et al , 2002 ), ( Ae ) HPV16 E6/E7-induced gene expression ( Nees et al , 2001 ), ( Af ) HPV31-induced gene expression , ( Ag ) Gene expression in SSC ( Loercher et al , 2004 ), ( Ah ) E2F-regulated genes ( Ishida et al , 2001 ), ( Ai ) HPV16- and HPV18-induced gene expression ( Santin et al , 2005 ) and, finally, ( Aj ) HPV18 E6/E7-induced gene expression ( Garner-Hamrick et al , 2004 ). ( B ) The total number of genes within each report cited. The numbers listed in parenthesis following the author are the total number with matching probes located on the ABI microarray. The number inside the large circle is the number of genes with matching probes in the 8449 selection. The number outside the big circle corresponds to genes with matching probes on the ABI chip but not included in the 8449 selection. ( C ) Hierarchical cluster analysis based on ( Ca ) CDKN2A (p16) transcriptionally regulated genes and ( Cb ) E2F-regulated genes, as reported by Vernell et al , 2003 .

Article Snippet: Immunohistochemistry was performed using a CDKN2A (p16)-specific antibody (BD Biosciences, Sparks, MD, USA, P/N 554070) diluted 1 : 200 in Antigen Diluent Solution, using a Dako Autostainer and the EnVision DAB detection system (Dako, P/N K5007).

Techniques: Selection, Expressing, Microarray

Journal: British Journal of Cancer

Article Title: Gene expression reveals two distinct groups of anal carcinomas with clinical implications

doi: 10.1038/sj.bjc.6604285

Figure Lengend Snippet: Immunohistochemical staining of MCM7 and CDKN2A proteins and ISH of HPV E7 mRNA is shown in ( A ), ( Aa ) MCM7 with predominant nuclear staining, ( Ab ) CDKN2A with nuclear and cytosolic staining and ( Ac ) ISH showing staining of predominantly nucleolar HPV E7 mRNA. ( B ) Immunohistochemical analysis of MCM7 protein expression in an anal carcinoma biopsy, illustrating one biopsy categorised as ‘high’ (upper pictures) ( Ba ) and one biopsy categorised as ‘low’ (lower pictures) ( Bb ) in the following Kaplan–Meier analysis. Kaplan–Meier estimates of RFS (upper graph) ( Bc ) and CSS (lower graph) ( Bd ) according to MCM7 protein expression among 55 patients with anal carcinomas.

Article Snippet: Immunohistochemistry was performed using a CDKN2A (p16)-specific antibody (BD Biosciences, Sparks, MD, USA, P/N 554070) diluted 1 : 200 in Antigen Diluent Solution, using a Dako Autostainer and the EnVision DAB detection system (Dako, P/N K5007).

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Discover Oncology

Article Title: Autophagy-related biomarkers in hepatocellular carcinoma and their relationship with immune infiltration

doi: 10.1007/s12672-024-01167-x

Figure Lengend Snippet: Lists of DEGs upregulated/downregulated in dataset GSE112790

Article Snippet: After blocking with a mixture of methanol and 0.75% hydrogen peroxide, sections were incubated with anti-IGF1 primary antibody (D160510, Sangon Biotech, China; dilution 1:50), anti-CDKN2A primary antibody (D227441, Sangon Biotech, China; dilution 1:65), anti-BIRC5 primary antibody (D120314, Sangon Biotech, China; dilution 1:60) and anti-SPP1 antibody (D121078, Sangon Biotech, China; dilution 1:20) at 4 °C overnight.

Techniques:

Journal: Discover Oncology

Article Title: Autophagy-related biomarkers in hepatocellular carcinoma and their relationship with immune infiltration

doi: 10.1007/s12672-024-01167-x

Figure Lengend Snippet: Information on differentially regulated genes

Article Snippet: After blocking with a mixture of methanol and 0.75% hydrogen peroxide, sections were incubated with anti-IGF1 primary antibody (D160510, Sangon Biotech, China; dilution 1:50), anti-CDKN2A primary antibody (D227441, Sangon Biotech, China; dilution 1:65), anti-BIRC5 primary antibody (D120314, Sangon Biotech, China; dilution 1:60) and anti-SPP1 antibody (D121078, Sangon Biotech, China; dilution 1:20) at 4 °C overnight.

Techniques:

Journal: Discover Oncology

Article Title: Autophagy-related biomarkers in hepatocellular carcinoma and their relationship with immune infiltration

doi: 10.1007/s12672-024-01167-x

Figure Lengend Snippet: Expression levels of IGF1, CDKN2A, BIRC 5, and SPP1 in HCC from the GEO and TCGA databases. A – H IGF1, CDKN2A, BIRC5, and SPP1 mRNA expression levels are based on the TCGA database, including paired ( A – D ) and nonpaired samples ( E – H ). I – L The mRNA expression levels of IGF1, CDKN2A, BIRC5, and SPP1 mRNA are based on GSE84402

Article Snippet: After blocking with a mixture of methanol and 0.75% hydrogen peroxide, sections were incubated with anti-IGF1 primary antibody (D160510, Sangon Biotech, China; dilution 1:50), anti-CDKN2A primary antibody (D227441, Sangon Biotech, China; dilution 1:65), anti-BIRC5 primary antibody (D120314, Sangon Biotech, China; dilution 1:60) and anti-SPP1 antibody (D121078, Sangon Biotech, China; dilution 1:20) at 4 °C overnight.

Techniques: Expressing

Journal: Discover Oncology

Article Title: Autophagy-related biomarkers in hepatocellular carcinoma and their relationship with immune infiltration

doi: 10.1007/s12672-024-01167-x

Figure Lengend Snippet: Levels of IGF1, CDKN2A, BIRC5, and SPP1 in normal individuals and patients with HCC from the HPA database

Article Snippet: After blocking with a mixture of methanol and 0.75% hydrogen peroxide, sections were incubated with anti-IGF1 primary antibody (D160510, Sangon Biotech, China; dilution 1:50), anti-CDKN2A primary antibody (D227441, Sangon Biotech, China; dilution 1:65), anti-BIRC5 primary antibody (D120314, Sangon Biotech, China; dilution 1:60) and anti-SPP1 antibody (D121078, Sangon Biotech, China; dilution 1:20) at 4 °C overnight.

Techniques:

Journal: Discover Oncology

Article Title: Autophagy-related biomarkers in hepatocellular carcinoma and their relationship with immune infiltration

doi: 10.1007/s12672-024-01167-x

Figure Lengend Snippet: Analysis of the clinical implications of IGF1, CDKN2A, BIRC5, and SPP1 on the XIANTAO planform. A – D ROC curve analysis for IGF1, CDKN2A, BIRC5, and SPP1. E – H Overall survival analysis for IGF1, CDKN2A, BIRC5, and SPP1. I – L Analysis of the association of TNM stages with IGF1, CDKN2A, BIRC5, and SPP1

Article Snippet: After blocking with a mixture of methanol and 0.75% hydrogen peroxide, sections were incubated with anti-IGF1 primary antibody (D160510, Sangon Biotech, China; dilution 1:50), anti-CDKN2A primary antibody (D227441, Sangon Biotech, China; dilution 1:65), anti-BIRC5 primary antibody (D120314, Sangon Biotech, China; dilution 1:60) and anti-SPP1 antibody (D121078, Sangon Biotech, China; dilution 1:20) at 4 °C overnight.

Techniques:

Journal: Discover Oncology

Article Title: Autophagy-related biomarkers in hepatocellular carcinoma and their relationship with immune infiltration

doi: 10.1007/s12672-024-01167-x

Figure Lengend Snippet: GSEA for IGF1, CDKN2A, BIRC5, and SPP1 by the TCGA database

Article Snippet: After blocking with a mixture of methanol and 0.75% hydrogen peroxide, sections were incubated with anti-IGF1 primary antibody (D160510, Sangon Biotech, China; dilution 1:50), anti-CDKN2A primary antibody (D227441, Sangon Biotech, China; dilution 1:65), anti-BIRC5 primary antibody (D120314, Sangon Biotech, China; dilution 1:60) and anti-SPP1 antibody (D121078, Sangon Biotech, China; dilution 1:20) at 4 °C overnight.

Techniques:

Journal: Discover Oncology

Article Title: Autophagy-related biomarkers in hepatocellular carcinoma and their relationship with immune infiltration

doi: 10.1007/s12672-024-01167-x

Figure Lengend Snippet: Levels of IGF1, CDKN2A, BIRC5, SPP1 and their correlations with the TME on the XIANTAO planform. A – D Immune infiltration analysis for IGF1, CDKN2A, BIRC5 and SPP1. E – H Correlation levels of IGF1, CDKN2A, BIRC5, and SPP1 with the level of immune infiltration

Article Snippet: After blocking with a mixture of methanol and 0.75% hydrogen peroxide, sections were incubated with anti-IGF1 primary antibody (D160510, Sangon Biotech, China; dilution 1:50), anti-CDKN2A primary antibody (D227441, Sangon Biotech, China; dilution 1:65), anti-BIRC5 primary antibody (D120314, Sangon Biotech, China; dilution 1:60) and anti-SPP1 antibody (D121078, Sangon Biotech, China; dilution 1:20) at 4 °C overnight.

Techniques:

Journal: Discover Oncology

Article Title: Autophagy-related biomarkers in hepatocellular carcinoma and their relationship with immune infiltration

doi: 10.1007/s12672-024-01167-x

Figure Lengend Snippet: Immune infiltration analysis of IGF1, CDKN2A, BIRC5, and SPP1 performed using the TIMER database

Article Snippet: After blocking with a mixture of methanol and 0.75% hydrogen peroxide, sections were incubated with anti-IGF1 primary antibody (D160510, Sangon Biotech, China; dilution 1:50), anti-CDKN2A primary antibody (D227441, Sangon Biotech, China; dilution 1:65), anti-BIRC5 primary antibody (D120314, Sangon Biotech, China; dilution 1:60) and anti-SPP1 antibody (D121078, Sangon Biotech, China; dilution 1:20) at 4 °C overnight.

Techniques:

Journal: Discover Oncology

Article Title: Autophagy-related biomarkers in hepatocellular carcinoma and their relationship with immune infiltration

doi: 10.1007/s12672-024-01167-x

Figure Lengend Snippet: Immune analysis of IGF1, CDKN2A, BIRC5, and SPP1 by the TCGA database. A – D Correlations of the estimated proportions of immune and stromal cells with IGF1, CDKN2A, BIRC5, and SPP1 levels in HCC. E Various immune checkpoints were associated with IGF1, CDKN2A, BIRC5, and SPP1

Article Snippet: After blocking with a mixture of methanol and 0.75% hydrogen peroxide, sections were incubated with anti-IGF1 primary antibody (D160510, Sangon Biotech, China; dilution 1:50), anti-CDKN2A primary antibody (D227441, Sangon Biotech, China; dilution 1:65), anti-BIRC5 primary antibody (D120314, Sangon Biotech, China; dilution 1:60) and anti-SPP1 antibody (D121078, Sangon Biotech, China; dilution 1:20) at 4 °C overnight.

Techniques:

Journal: Discover Oncology

Article Title: Autophagy-related biomarkers in hepatocellular carcinoma and their relationship with immune infiltration

doi: 10.1007/s12672-024-01167-x

Figure Lengend Snippet: Levels of IGF1, CDKN2A, BIRC5, and SPP1 in patient tissues by IHC. Levels of CDKN2A, BIRC5, and SPP1 in HCC were increased compared with tumor-adjacent tissues. However, IGF1 expression was downregulated in HCC tissues compared with adjacent tissues

Article Snippet: After blocking with a mixture of methanol and 0.75% hydrogen peroxide, sections were incubated with anti-IGF1 primary antibody (D160510, Sangon Biotech, China; dilution 1:50), anti-CDKN2A primary antibody (D227441, Sangon Biotech, China; dilution 1:65), anti-BIRC5 primary antibody (D120314, Sangon Biotech, China; dilution 1:60) and anti-SPP1 antibody (D121078, Sangon Biotech, China; dilution 1:20) at 4 °C overnight.

Techniques: Expressing

Journal: Discover Oncology

Article Title: Autophagy-related biomarkers in hepatocellular carcinoma and their relationship with immune infiltration

doi: 10.1007/s12672-024-01167-x

Figure Lengend Snippet: Expressions of IGF1, CDKN2A, BIRC5, and SPP1 in patient tissues by RT‒PCR. The expressions of CDKN2A, SPP1, and BIRC5 in HCC were increased compared with tumor-adjacent tissues. However, IGF1 expression was downregulated in HCC tissues compared with that in adjacent tissues

Article Snippet: After blocking with a mixture of methanol and 0.75% hydrogen peroxide, sections were incubated with anti-IGF1 primary antibody (D160510, Sangon Biotech, China; dilution 1:50), anti-CDKN2A primary antibody (D227441, Sangon Biotech, China; dilution 1:65), anti-BIRC5 primary antibody (D120314, Sangon Biotech, China; dilution 1:60) and anti-SPP1 antibody (D121078, Sangon Biotech, China; dilution 1:20) at 4 °C overnight.

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: rhCC16 Suppresses Cellular Senescence and Ameliorates COPD‐Like Symptoms by Activating the AMPK/Sirt1‐PGC‐1‐α‐TFAM Pathway to Promote Mitochondrial Function

doi: 10.1111/jcmm.70566

Figure Lengend Snippet: Primers sets used for real‐time PCR.

Article Snippet: The primary and secondary antibodies used in the present study were as follows: anti‐GAPDH, anti‐PARP and anti‐Uteroglobin (CC16) antibodies purchased from Bioss (Beijing, China); anti‐CDKN2A/p16 INK4α , anti‐CDKN1A/p21 CIP1 , anti‐Caspase3 and anti‐GLB1 antibodies purchased from ABclonal (Wuhan, China); anti‐NDUFB10, anti‐SDHA, and anti‐UQCRC2 antibodies purchased from Sangon Biotechnology (Shanghai, China); anti‐AMPK, anti‐p‐AMPK, anti‐Sirt1, anti‐PGC‐1α, and anti‐TFAM antibodies purchased from Beyotime Biotechnology (Shanghai, China); and HRP‐labelled goat anti‐rabbit/mouse IgG (H + L) purchased from Boster Biotechnology (Wuhan, China).

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: rhCC16 Suppresses Cellular Senescence and Ameliorates COPD‐Like Symptoms by Activating the AMPK/Sirt1‐PGC‐1‐α‐TFAM Pathway to Promote Mitochondrial Function

doi: 10.1111/jcmm.70566

Figure Lengend Snippet: Treatment with rhCC16 ameliorated cellular senescence in mouse lung tissue cells and COPD‐like pathological changes in mice with COPD. (A) The levels of CC16 protein in the lung tissues of mice at different ages were measured using ELISA. (B) Representative immunohistochemical images of CC16 and GLB1 in the lung tissues of mice; scale bar: 100 μm. (C) The statistical graph of the data obtained from the immunohistochemistry analysis presented in (B) were expressed as positive cell area fractions. (D) Results of lung function tests conducted with mice at different ages. (E) Representative images of P16, P21, and GLB1 obtained by immunohistochemical analysis in the control, COPD, and rhCC16 groups; scale bar: 200 μm. (F) Representative images of H&E‐stained mouse lung tissue samples from the control, COPD, and rhCC16 groups. The mice in the COPD group and rhCC16 group were exposed to smoke for 24 weeks, 5 days a week, for 2 h each day. In the 21st week, the mice in the rhCC16 group were treated with intranasal administration 2 h before smoke exposure, and the treatment lasted for 1 month. After 24 weeks, fresh lung tissues were collected, embedded, and paraffin sections were prepared for HE staining. The data were presented as the mean ± SEM. * p < 0.05 and ** p < 0.01, *** p < 0.001, compared to the control group; # p < 0.05, ## p < 0.01, compared to the COPD group.

Article Snippet: The primary and secondary antibodies used in the present study were as follows: anti‐GAPDH, anti‐PARP and anti‐Uteroglobin (CC16) antibodies purchased from Bioss (Beijing, China); anti‐CDKN2A/p16 INK4α , anti‐CDKN1A/p21 CIP1 , anti‐Caspase3 and anti‐GLB1 antibodies purchased from ABclonal (Wuhan, China); anti‐NDUFB10, anti‐SDHA, and anti‐UQCRC2 antibodies purchased from Sangon Biotechnology (Shanghai, China); anti‐AMPK, anti‐p‐AMPK, anti‐Sirt1, anti‐PGC‐1α, and anti‐TFAM antibodies purchased from Beyotime Biotechnology (Shanghai, China); and HRP‐labelled goat anti‐rabbit/mouse IgG (H + L) purchased from Boster Biotechnology (Wuhan, China).

Techniques: Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Immunohistochemistry, Control, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: rhCC16 Suppresses Cellular Senescence and Ameliorates COPD‐Like Symptoms by Activating the AMPK/Sirt1‐PGC‐1‐α‐TFAM Pathway to Promote Mitochondrial Function

doi: 10.1111/jcmm.70566

Figure Lengend Snippet: rhCC16 inhibited the senescence induced by CSE. (A) The protein levels of P16 and P21 in CSE‐stimulated BEAS‐2B cells were determined using Western blot. (B) The mRNA expression of P16 and P21 in CSE‐stimulated BEAS‐2B cells was determined through RT–qPCR. (C) Representative images of the senescence‐associated β‐galactosidase staining experiment conducted with CSE‐stimulated BEAS‐2B cells; scale bar: 20 μm. (D) The protein levels of P16 and P21 in CSE‐stimulated or rhCC16‐treated BEAS‐2B cells were determined using Western blot. (E) The mRNA expression of P16 and P21 in CSE‐stimulated or rhCC16‐treated BEAS‐2B cells was determined through RT–qPCR. (F) Representative images of senescence‐associated β‐galactosidase staining in CSE‐stimulated or rhCC16‐treated BEAS‐2B cells; scale bar: 20 μm. The data were presented as the mean ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001, compared to the control group; # p < 0.05, ## p < 0.01, compared to the 5% CSE group.

Article Snippet: The primary and secondary antibodies used in the present study were as follows: anti‐GAPDH, anti‐PARP and anti‐Uteroglobin (CC16) antibodies purchased from Bioss (Beijing, China); anti‐CDKN2A/p16 INK4α , anti‐CDKN1A/p21 CIP1 , anti‐Caspase3 and anti‐GLB1 antibodies purchased from ABclonal (Wuhan, China); anti‐NDUFB10, anti‐SDHA, and anti‐UQCRC2 antibodies purchased from Sangon Biotechnology (Shanghai, China); anti‐AMPK, anti‐p‐AMPK, anti‐Sirt1, anti‐PGC‐1α, and anti‐TFAM antibodies purchased from Beyotime Biotechnology (Shanghai, China); and HRP‐labelled goat anti‐rabbit/mouse IgG (H + L) purchased from Boster Biotechnology (Wuhan, China).

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Staining, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: rhCC16 Suppresses Cellular Senescence and Ameliorates COPD‐Like Symptoms by Activating the AMPK/Sirt1‐PGC‐1‐α‐TFAM Pathway to Promote Mitochondrial Function

doi: 10.1111/jcmm.70566

Figure Lengend Snippet: rhCC16 improved mitochondrial function via the AMPK/Sirt1/PGC‐1α/TFAM pathway both in vitro and in vivo. (A) Representative images of the oxidative phosphorylation complex I, complex II, and complex III proteins obtained via immunohistochemistry analysis of mouse lung tissue samples from the control, COPD, and rhCC16 treatment groups; scale bar: 200 μm. (B) Representative images of p‐AMPK, SIRT1, PGC‐1α, and TFAM obtained from immunohistochemical analysis of mouse lung tissue samples from the control, COPD, and rhCC16 treatment groups; scale bar: 200 μm. (C) Statistical bar chart plot for Figure A and Figure B. (D) Representative TEM images of mouse lung tissue samples from the control, COPD, and rhCC16 treatment groups; yellow arrows represent mitochondria with normal morphology; red arrows represent mitochondria with abnormal morphology and functional damage; blue arrows represent mitochondria with restored morphology and function. Scale bar: 500 nm. (E) The levels of p16 and p21 proteins in CSE‐stimulated or rhCC16‐treated BEAS‐2B cells treated with BIO‐1211 (10 μM) or M‐β‐CD (5 μM) were determined using Western blot. The data were presented as the mean ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001, compared to the control group; # p < 0.05, ## p < 0.01, and ### p < 0.001, compared to the 5% CSE group; & p < 0.05, && p < 0.01, compared to the 5% CSE + rhCC16 group.

Article Snippet: The primary and secondary antibodies used in the present study were as follows: anti‐GAPDH, anti‐PARP and anti‐Uteroglobin (CC16) antibodies purchased from Bioss (Beijing, China); anti‐CDKN2A/p16 INK4α , anti‐CDKN1A/p21 CIP1 , anti‐Caspase3 and anti‐GLB1 antibodies purchased from ABclonal (Wuhan, China); anti‐NDUFB10, anti‐SDHA, and anti‐UQCRC2 antibodies purchased from Sangon Biotechnology (Shanghai, China); anti‐AMPK, anti‐p‐AMPK, anti‐Sirt1, anti‐PGC‐1α, and anti‐TFAM antibodies purchased from Beyotime Biotechnology (Shanghai, China); and HRP‐labelled goat anti‐rabbit/mouse IgG (H + L) purchased from Boster Biotechnology (Wuhan, China).

Techniques: In Vitro, In Vivo, Phospho-proteomics, Immunohistochemistry, Control, Immunohistochemical staining, Functional Assay, Western Blot