anti oc2  (Proteintech)

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Knockout of onecut2 inhibits proliferation and promotes apoptosis of tumor cells through SKP2-mediated p53 acetylation in hepatocellular carcinoma

doi: 10.1007/s00018-024-05518-3

Figure Lengend Snippet: OC2 knockdown inhibited proliferation and promoted apoptosis in HCC cells. A - B The expression of OC2 in HCC clinical samples was analyzed by qRT‒PCR and Western blotting. “T” represents tumor tissue, and “N” represents normal tissue. C The expression of OC2 in HCC clinical tissues was analyzed by IHC. Scale bar: 200 μm. D - E The expression of OC2 in normal cells (HEK293T and HUVEC) and HCC cells (SK-Hep1, PLC/PRF/5, Huh6 and HepG2) was analyzed by qRT‒PCR and Western blotting. F - G Results of qRT‒PCR and Western blot analysis of the effects of OC2 interference by shRNA (OC2 knockdown) in HCC cells. H - I Results of the CCK-8 and plate colony formation assays with OC2 knockdown in HCC cells. J The phosphorylation levels of AKT and MAPK and the expression of FGF2 in HCC cells were analyzed by Western blotting. K The apoptosis of HCC cells with OC2 knockdown was analyzed by flow cytometry. L - M The expression of apoptosis-related factors (PARP, Bcl2 and Bax) in HCC cells was analyzed by qRT‒PCR and Western blotting. The data are expressed as mean ± SD; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The primary antibodies used included an anti-OC2 antibody purchased from Proteintech (21916-1-AP).

Techniques: Knockdown, Expressing, Western Blot, shRNA, CCK-8 Assay, Phospho-proteomics, Flow Cytometry

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Knockout of onecut2 inhibits proliferation and promotes apoptosis of tumor cells through SKP2-mediated p53 acetylation in hepatocellular carcinoma

doi: 10.1007/s00018-024-05518-3

Figure Lengend Snippet: OC2 knockout inhibited proliferation and migration and promoted apoptosis. A Recognition sites of sgRNA1 and sgRNA2 in exon 1 of the OC2 gene. B - C The relative expression of OC2 in B8 and B10 cells was analyzed by Western blotting and qRT‒PCR. The “blank” group consisted of SK-Hep1 cells that were not transfected with lentivirus; the “Con” (Control) group consisted of SK-Hep1 cells that were transfected with lentivirus without a knockout site; the “B8” and “B10” groups consisted of SK-Hep1 cells that were transfected with lentivirus containing the sgRNA1 and sgRNA2 knockout sites. D - E The proliferation of B8 and B10 cells was analyzed by CCK-8 and plate colony formation assays. F The expression of ATK, MAPK, and FGF2 in B8 and B10 cells was analyzed by qRT‒PCR. G The phosphorylation levels of ATK and MAPK and the expression of FGF2 in B8 and B10 cells was analyzed by Western blotting. H Tube formation assay of HUVECs treated with the supernatant of B8 and B10 cells. Scale bar: 100 μm. I - J The expression of angiogenesis-related factors (CD31, PDGF-B, VEGFA, angiogenin and TGF-β1) in B8 and B10 cells was analyzed by qRT‒PCR and Western blotting. K Apoptosis results from the flow cytometry assay in B8 and B10 cells. L - M The expression of apoptosis-related factors in B8 and B10 cells was analyzed by qRT‒PCR and Western blotting. The data are expressed as mean ± SD; “ns” indicates no significant difference; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The primary antibodies used included an anti-OC2 antibody purchased from Proteintech (21916-1-AP).

Techniques: Knock-Out, Migration, Expressing, Western Blot, Transfection, Control, CCK-8 Assay, Phospho-proteomics, Tube Formation Assay, Flow Cytometry

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Knockout of onecut2 inhibits proliferation and promotes apoptosis of tumor cells through SKP2-mediated p53 acetylation in hepatocellular carcinoma

doi: 10.1007/s00018-024-05518-3

Figure Lengend Snippet: Identification of OC2-binding genes via ChIP‒Seq, qRT‒PCR and ChIP‒qPCR. A - B The relative expression of candidate genes analyzed by qRT‒PCR after OC2 was overexpressed in HepG2 and SK-Hep1 cells. The “OENC” group consisted of SK-Hep1 or HepG2 cells transfected with overexpression empty plasmid; The “OEOC2” group consisted of SK-Hep1 or HepG2 cells transfected with OC2-overexpressing plasmid; C The relative expression of candidate genes analyzed by qRT‒PCR in B8 cells. The “NC” group consisted of SK-Hep1 cells that were transfected with lentivirus without a knockout site; the “B8” group consisted of SK-Hep1 cells with OC2 knockout. D The promoter activities of DSP and SKP2 were analyzed via dual-luciferase reporter assays. The “pGL3-Basic” group consisted of SK-Hep1 cells transfected with an empty plasmid without the promoter sequence inserted; The “pGL3-DSP” group consisted of SK-Hep1 cells transfected with a plasmid inserted with the DSP promoter sequence; The “pGL3-SKP2” group consisted of SK-Hep1 cells transfected with a plasmid inserted with the SKP2 promoter sequence. E Motif of the OC2 transcription factor binding to DNA. F Mutations in the OC2 binding site in the SKP2 promoter sequence. The bases marked in red are the base sequences after mutation. G The mutant promoter activity of SKP2 was analyzed via dual-luciferase reporter assays. The “pGL3-SKP2-WT” group consisted of SK-Hep1 cells transfected with a plasmid inserted with the normal SKP2 promoter sequence; The “pGL3-SKP2-mut” group consisted of SK-Hep1 cells transfected with a plasmid inserted with the mutated SKP2 promoter sequence. H - I The expression of SKP2 in HCC clinical samples was analyzed by Western blotting and qRT‒PCR. J Correlation analysis of the relative expression levels of OC2 and SKP2 in HCC clinical samples. K The relative expression of SKP2 after OC2 knockdown in HepG2 and SK-Hep1 cells was analyzed by Western blotting. L Relative fold enrichment of SKP2 promoter fragments analyzed by ChIP‒qPCR in HepG2 and SK-Hep1 cells. The “IgG” group was immunoprecipitated using anti-IgG antibody; the “OENC” and “OEOC2” groups were immunoprecipitated using anti-OC2 antibody. The data are expressed as mean ± SD; “ns” indicates no significant difference; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The primary antibodies used included an anti-OC2 antibody purchased from Proteintech (21916-1-AP).

Techniques: Binding Assay, Expressing, Transfection, Over Expression, Plasmid Preparation, Knock-Out, Luciferase, Sequencing, Mutagenesis, Activity Assay, Western Blot, Knockdown, Immunoprecipitation

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Knockout of onecut2 inhibits proliferation and promotes apoptosis of tumor cells through SKP2-mediated p53 acetylation in hepatocellular carcinoma

doi: 10.1007/s00018-024-05518-3

Figure Lengend Snippet: SKP2 knockdown inhibited proliferation and promoted apoptosis in HCC cells. A - B SKP2 knockdown with shRNA in HepG2 and SK-Hep1 cells was analyzed by qRT‒PCR and Western blotting. C SKP2 knockdown inhibited the proliferation of HepG2 and SK-Hep1 cells, as determined by CCK-8 assays. D SKP2 knockdown promoted the apoptosis of HepG2 and SK-Hep1 cells, as determined by flow cytometry. E - F SKP2 knockdown increased the expression of apoptosis-related factors, as determined by qRT‒PCR and Western blotting, in HepG2 and SK-Hep1 cells. G - H SKP2 knockdown promoted the expression of p21 and p27, as determined by qRT‒PCR and Western blotting, in HepG2 and SK-Hep1 cells. I OC2 knockdown increased the expression of p21 and p27, as determined by Western blotting, in HepG2 and SK-Hep1 cells. The data are expressed as mean ± SD; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The primary antibodies used included an anti-OC2 antibody purchased from Proteintech (21916-1-AP).

Techniques: Knockdown, shRNA, Western Blot, CCK-8 Assay, Flow Cytometry, Expressing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Knockout of onecut2 inhibits proliferation and promotes apoptosis of tumor cells through SKP2-mediated p53 acetylation in hepatocellular carcinoma

doi: 10.1007/s00018-024-05518-3

Figure Lengend Snippet: Role of proliferation and apoptosis mediated by the overexpression of OC2 and SKP2 in B8 cells. A The proliferation of B8 cells overexpressing OC2 was analyzed via a CCK-8 assay. B Flow cytometry assay of apoptosis levels in B8 cells overexpressing OC2. C The expression of apoptosis-related factors (SKP2, PARP, Bax, Bcl2, p21 and p27) in B8 cells overexpressing OC2. D Proliferation of B8 cells overexpressing SKP2. E Flow cytometry assay of apoptosis levels in B8 cells overexpressing SKP2. F - G The expression of apoptosis-related factors in B8 cells overexpressing SKP2 was analyzed by Western blotting and qRT‒PCR. H The phosphorylation and acetylation levels of p53 were analyzed by Western blotting in B8 cells overexpressing SKP2. I Dual-luciferase reporter assays of p53 response element-related fluorescence activity in SK-Hep1 cells with OC2 and SKP2 knockdown. The “p53-RE” group consisted of SK-Hep1 cells transfected with pGPL3-Basic plasmid with p53 response element sequence inserted. J Phosphorylation and acetylation levels of p53 in B8 cells overexpressing OC2 and SKP2. The data are expressed as mean ± SD; “ns” indicates no significant difference; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The primary antibodies used included an anti-OC2 antibody purchased from Proteintech (21916-1-AP).

Techniques: Over Expression, CCK-8 Assay, Flow Cytometry, Expressing, Western Blot, Phospho-proteomics, Luciferase, Fluorescence, Activity Assay, Knockdown, Transfection, Plasmid Preparation, Sequencing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Knockout of onecut2 inhibits proliferation and promotes apoptosis of tumor cells through SKP2-mediated p53 acetylation in hepatocellular carcinoma

doi: 10.1007/s00018-024-05518-3

Figure Lengend Snippet: Effect of OC2 knockout on tumor growth in vivo. A Schematic diagram of the xenograft mouse model. The volume of the tumor was measured every three days after the tumor was subcutaneously formed in the mouse. B Tumor tissues were removed from the mice in the B8 and B10 cell groups and the control group. C - D Tumor growth curves and tumor weights. E The phosphorylation levels of AKT and MAPK and the expression of FGF2 in tumor tissues were analyzed by Western blotting. F The expression of angiogenesis-related factors in tumor tissues was analyzed by Western blotting. G The expression of apoptosis-related factors in tumor tissues was analyzed by Western blotting. H Results of immunohistochemistry and TUNEL assays for tumor growth and apoptosis. Scale bar: 100 μm; the data are expressed as mean ± SD; *** P < 0.001

Article Snippet: The primary antibodies used included an anti-OC2 antibody purchased from Proteintech (21916-1-AP).

Techniques: Knock-Out, In Vivo, Control, Phospho-proteomics, Expressing, Western Blot, Immunohistochemistry, TUNEL Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Knockout of onecut2 inhibits proliferation and promotes apoptosis of tumor cells through SKP2-mediated p53 acetylation in hepatocellular carcinoma

doi: 10.1007/s00018-024-05518-3

Figure Lengend Snippet: The mechanism by which OC2 regulates proliferation and apoptosis in HCC cells. OC2 promoted the transcriptional activation of SKP2, which inhibited the binding of p53 to p300 and downregulated the acetylation level of p53, thereby inhibiting the apoptosis of HCC cells. The solid line indicates direct action; the dotted line indicates indirect action; the “AC” indicates acetylation; the “P” indicates phosphorylation

Article Snippet: The primary antibodies used included an anti-OC2 antibody purchased from Proteintech (21916-1-AP).

Techniques: Activation Assay, Binding Assay, Phospho-proteomics

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Knockout of onecut2 inhibits proliferation and promotes apoptosis of tumor cells through SKP2-mediated p53 acetylation in hepatocellular carcinoma

doi: 10.1007/s00018-024-05518-3

Figure Lengend Snippet: OC2 knockdown inhibited proliferation and promoted apoptosis in HCC cells. A - B The expression of OC2 in HCC clinical samples was analyzed by qRT‒PCR and Western blotting. “T” represents tumor tissue, and “N” represents normal tissue. C The expression of OC2 in HCC clinical tissues was analyzed by IHC. Scale bar: 200 μm. D - E The expression of OC2 in normal cells (HEK293T and HUVEC) and HCC cells (SK-Hep1, PLC/PRF/5, Huh6 and HepG2) was analyzed by qRT‒PCR and Western blotting. F - G Results of qRT‒PCR and Western blot analysis of the effects of OC2 interference by shRNA (OC2 knockdown) in HCC cells. H - I Results of the CCK-8 and plate colony formation assays with OC2 knockdown in HCC cells. J The phosphorylation levels of AKT and MAPK and the expression of FGF2 in HCC cells were analyzed by Western blotting. K The apoptosis of HCC cells with OC2 knockdown was analyzed by flow cytometry. L - M The expression of apoptosis-related factors (PARP, Bcl2 and Bax) in HCC cells was analyzed by qRT‒PCR and Western blotting. The data are expressed as mean ± SD; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The sections were incubated with primary antibodies against OC2 (Proteintech, 21916-1-AP, 1:250), SKP2 (Cell Signaling Technology, 2652, 1:200), KI67 (Cell Signaling Technology, 12202, 1:400) and CD31 (Cell Signaling Technology, 77699, 1:200) at 4 °C overnight.

Techniques: Knockdown, Expressing, Western Blot, shRNA, CCK-8 Assay, Phospho-proteomics, Flow Cytometry

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Knockout of onecut2 inhibits proliferation and promotes apoptosis of tumor cells through SKP2-mediated p53 acetylation in hepatocellular carcinoma

doi: 10.1007/s00018-024-05518-3

Figure Lengend Snippet: OC2 knockout inhibited proliferation and migration and promoted apoptosis. A Recognition sites of sgRNA1 and sgRNA2 in exon 1 of the OC2 gene. B - C The relative expression of OC2 in B8 and B10 cells was analyzed by Western blotting and qRT‒PCR. The “blank” group consisted of SK-Hep1 cells that were not transfected with lentivirus; the “Con” (Control) group consisted of SK-Hep1 cells that were transfected with lentivirus without a knockout site; the “B8” and “B10” groups consisted of SK-Hep1 cells that were transfected with lentivirus containing the sgRNA1 and sgRNA2 knockout sites. D - E The proliferation of B8 and B10 cells was analyzed by CCK-8 and plate colony formation assays. F The expression of ATK, MAPK, and FGF2 in B8 and B10 cells was analyzed by qRT‒PCR. G The phosphorylation levels of ATK and MAPK and the expression of FGF2 in B8 and B10 cells was analyzed by Western blotting. H Tube formation assay of HUVECs treated with the supernatant of B8 and B10 cells. Scale bar: 100 μm. I - J The expression of angiogenesis-related factors (CD31, PDGF-B, VEGFA, angiogenin and TGF-β1) in B8 and B10 cells was analyzed by qRT‒PCR and Western blotting. K Apoptosis results from the flow cytometry assay in B8 and B10 cells. L - M The expression of apoptosis-related factors in B8 and B10 cells was analyzed by qRT‒PCR and Western blotting. The data are expressed as mean ± SD; “ns” indicates no significant difference; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The sections were incubated with primary antibodies against OC2 (Proteintech, 21916-1-AP, 1:250), SKP2 (Cell Signaling Technology, 2652, 1:200), KI67 (Cell Signaling Technology, 12202, 1:400) and CD31 (Cell Signaling Technology, 77699, 1:200) at 4 °C overnight.

Techniques: Knock-Out, Migration, Expressing, Western Blot, Transfection, Control, CCK-8 Assay, Phospho-proteomics, Tube Formation Assay, Flow Cytometry

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Knockout of onecut2 inhibits proliferation and promotes apoptosis of tumor cells through SKP2-mediated p53 acetylation in hepatocellular carcinoma

doi: 10.1007/s00018-024-05518-3

Figure Lengend Snippet: Identification of OC2-binding genes via ChIP‒Seq, qRT‒PCR and ChIP‒qPCR. A - B The relative expression of candidate genes analyzed by qRT‒PCR after OC2 was overexpressed in HepG2 and SK-Hep1 cells. The “OENC” group consisted of SK-Hep1 or HepG2 cells transfected with overexpression empty plasmid; The “OEOC2” group consisted of SK-Hep1 or HepG2 cells transfected with OC2-overexpressing plasmid; C The relative expression of candidate genes analyzed by qRT‒PCR in B8 cells. The “NC” group consisted of SK-Hep1 cells that were transfected with lentivirus without a knockout site; the “B8” group consisted of SK-Hep1 cells with OC2 knockout. D The promoter activities of DSP and SKP2 were analyzed via dual-luciferase reporter assays. The “pGL3-Basic” group consisted of SK-Hep1 cells transfected with an empty plasmid without the promoter sequence inserted; The “pGL3-DSP” group consisted of SK-Hep1 cells transfected with a plasmid inserted with the DSP promoter sequence; The “pGL3-SKP2” group consisted of SK-Hep1 cells transfected with a plasmid inserted with the SKP2 promoter sequence. E Motif of the OC2 transcription factor binding to DNA. F Mutations in the OC2 binding site in the SKP2 promoter sequence. The bases marked in red are the base sequences after mutation. G The mutant promoter activity of SKP2 was analyzed via dual-luciferase reporter assays. The “pGL3-SKP2-WT” group consisted of SK-Hep1 cells transfected with a plasmid inserted with the normal SKP2 promoter sequence; The “pGL3-SKP2-mut” group consisted of SK-Hep1 cells transfected with a plasmid inserted with the mutated SKP2 promoter sequence. H - I The expression of SKP2 in HCC clinical samples was analyzed by Western blotting and qRT‒PCR. J Correlation analysis of the relative expression levels of OC2 and SKP2 in HCC clinical samples. K The relative expression of SKP2 after OC2 knockdown in HepG2 and SK-Hep1 cells was analyzed by Western blotting. L Relative fold enrichment of SKP2 promoter fragments analyzed by ChIP‒qPCR in HepG2 and SK-Hep1 cells. The “IgG” group was immunoprecipitated using anti-IgG antibody; the “OENC” and “OEOC2” groups were immunoprecipitated using anti-OC2 antibody. The data are expressed as mean ± SD; “ns” indicates no significant difference; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The sections were incubated with primary antibodies against OC2 (Proteintech, 21916-1-AP, 1:250), SKP2 (Cell Signaling Technology, 2652, 1:200), KI67 (Cell Signaling Technology, 12202, 1:400) and CD31 (Cell Signaling Technology, 77699, 1:200) at 4 °C overnight.

Techniques: Binding Assay, Expressing, Transfection, Over Expression, Plasmid Preparation, Knock-Out, Luciferase, Sequencing, Mutagenesis, Activity Assay, Western Blot, Knockdown, Immunoprecipitation

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Knockout of onecut2 inhibits proliferation and promotes apoptosis of tumor cells through SKP2-mediated p53 acetylation in hepatocellular carcinoma

doi: 10.1007/s00018-024-05518-3

Figure Lengend Snippet: SKP2 knockdown inhibited proliferation and promoted apoptosis in HCC cells. A - B SKP2 knockdown with shRNA in HepG2 and SK-Hep1 cells was analyzed by qRT‒PCR and Western blotting. C SKP2 knockdown inhibited the proliferation of HepG2 and SK-Hep1 cells, as determined by CCK-8 assays. D SKP2 knockdown promoted the apoptosis of HepG2 and SK-Hep1 cells, as determined by flow cytometry. E - F SKP2 knockdown increased the expression of apoptosis-related factors, as determined by qRT‒PCR and Western blotting, in HepG2 and SK-Hep1 cells. G - H SKP2 knockdown promoted the expression of p21 and p27, as determined by qRT‒PCR and Western blotting, in HepG2 and SK-Hep1 cells. I OC2 knockdown increased the expression of p21 and p27, as determined by Western blotting, in HepG2 and SK-Hep1 cells. The data are expressed as mean ± SD; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The sections were incubated with primary antibodies against OC2 (Proteintech, 21916-1-AP, 1:250), SKP2 (Cell Signaling Technology, 2652, 1:200), KI67 (Cell Signaling Technology, 12202, 1:400) and CD31 (Cell Signaling Technology, 77699, 1:200) at 4 °C overnight.

Techniques: Knockdown, shRNA, Western Blot, CCK-8 Assay, Flow Cytometry, Expressing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Knockout of onecut2 inhibits proliferation and promotes apoptosis of tumor cells through SKP2-mediated p53 acetylation in hepatocellular carcinoma

doi: 10.1007/s00018-024-05518-3

Figure Lengend Snippet: Role of proliferation and apoptosis mediated by the overexpression of OC2 and SKP2 in B8 cells. A The proliferation of B8 cells overexpressing OC2 was analyzed via a CCK-8 assay. B Flow cytometry assay of apoptosis levels in B8 cells overexpressing OC2. C The expression of apoptosis-related factors (SKP2, PARP, Bax, Bcl2, p21 and p27) in B8 cells overexpressing OC2. D Proliferation of B8 cells overexpressing SKP2. E Flow cytometry assay of apoptosis levels in B8 cells overexpressing SKP2. F - G The expression of apoptosis-related factors in B8 cells overexpressing SKP2 was analyzed by Western blotting and qRT‒PCR. H The phosphorylation and acetylation levels of p53 were analyzed by Western blotting in B8 cells overexpressing SKP2. I Dual-luciferase reporter assays of p53 response element-related fluorescence activity in SK-Hep1 cells with OC2 and SKP2 knockdown. The “p53-RE” group consisted of SK-Hep1 cells transfected with pGPL3-Basic plasmid with p53 response element sequence inserted. J Phosphorylation and acetylation levels of p53 in B8 cells overexpressing OC2 and SKP2. The data are expressed as mean ± SD; “ns” indicates no significant difference; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The sections were incubated with primary antibodies against OC2 (Proteintech, 21916-1-AP, 1:250), SKP2 (Cell Signaling Technology, 2652, 1:200), KI67 (Cell Signaling Technology, 12202, 1:400) and CD31 (Cell Signaling Technology, 77699, 1:200) at 4 °C overnight.

Techniques: Over Expression, CCK-8 Assay, Flow Cytometry, Expressing, Western Blot, Phospho-proteomics, Luciferase, Fluorescence, Activity Assay, Knockdown, Transfection, Plasmid Preparation, Sequencing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Knockout of onecut2 inhibits proliferation and promotes apoptosis of tumor cells through SKP2-mediated p53 acetylation in hepatocellular carcinoma

doi: 10.1007/s00018-024-05518-3

Figure Lengend Snippet: Effect of OC2 knockout on tumor growth in vivo. A Schematic diagram of the xenograft mouse model. The volume of the tumor was measured every three days after the tumor was subcutaneously formed in the mouse. B Tumor tissues were removed from the mice in the B8 and B10 cell groups and the control group. C - D Tumor growth curves and tumor weights. E The phosphorylation levels of AKT and MAPK and the expression of FGF2 in tumor tissues were analyzed by Western blotting. F The expression of angiogenesis-related factors in tumor tissues was analyzed by Western blotting. G The expression of apoptosis-related factors in tumor tissues was analyzed by Western blotting. H Results of immunohistochemistry and TUNEL assays for tumor growth and apoptosis. Scale bar: 100 μm; the data are expressed as mean ± SD; *** P < 0.001

Article Snippet: The sections were incubated with primary antibodies against OC2 (Proteintech, 21916-1-AP, 1:250), SKP2 (Cell Signaling Technology, 2652, 1:200), KI67 (Cell Signaling Technology, 12202, 1:400) and CD31 (Cell Signaling Technology, 77699, 1:200) at 4 °C overnight.

Techniques: Knock-Out, In Vivo, Control, Phospho-proteomics, Expressing, Western Blot, Immunohistochemistry, TUNEL Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Knockout of onecut2 inhibits proliferation and promotes apoptosis of tumor cells through SKP2-mediated p53 acetylation in hepatocellular carcinoma

doi: 10.1007/s00018-024-05518-3

Figure Lengend Snippet: The mechanism by which OC2 regulates proliferation and apoptosis in HCC cells. OC2 promoted the transcriptional activation of SKP2, which inhibited the binding of p53 to p300 and downregulated the acetylation level of p53, thereby inhibiting the apoptosis of HCC cells. The solid line indicates direct action; the dotted line indicates indirect action; the “AC” indicates acetylation; the “P” indicates phosphorylation

Article Snippet: The sections were incubated with primary antibodies against OC2 (Proteintech, 21916-1-AP, 1:250), SKP2 (Cell Signaling Technology, 2652, 1:200), KI67 (Cell Signaling Technology, 12202, 1:400) and CD31 (Cell Signaling Technology, 77699, 1:200) at 4 °C overnight.

Techniques: Activation Assay, Binding Assay, Phospho-proteomics

Journal: Frontiers in Pharmacology

Article Title: Hyperglycemia-independent neonatal streptozotocin-induced retinopathy (NSIR) in rats

doi: 10.3389/fphar.2024.1395887

Figure Lengend Snippet: Intravitreal STZ injection on P1 inhibits the proliferation of neonatal rat retinal progenitors and delays their cell cycle exit and differentiation. (A) Horizontal P4 retinal sections of indicated locations and groups were stained for nuclear (DAPI, blue) and cell proliferation (Ki67, green). (B) Quantification of Ki67+ cells per mm 2 area of P4 and P8 retinas. (C) Horizontal P8 retinal sections of indicated locations and groups were stained for nuclear (DAPI, blue) and cell proliferation (Ki67, green). (D) Flow cytometry analysis of P4 retinal cells. (E) Whole-mount or horizontal sections of P8 retinas of indicated groups were stained for nuclear (DAPI, blue) and cell type markers, including Ganglion cells (Brn3, green, whole-mount), Cone (ARR3, green), Horizontal cells (OC2, green), Amacrine cells (Ap2a, green), Rod (Rho, green), Bipolar cells (Chx10, green) and Müller glia (Sox9, green). (F) Quantifying all seven retinal cell types (relative to the control group, %). Error bars represent SD of measurements from three animals or three retinas of three animals (n = 3), and asterisks indicate significant differences between control and STZ-treated groups (* p < 0.05, ** p < 0.01, one-way ANOVA followed by Bonferroni’s correction). ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer; Ivit, Intravitreal injection; PI, Propidium iodide. Scale bar: 50 μm.

Article Snippet: Slides were incubated with blocking solution (1% donkey serum and 0.1% Triton X-100 in PBS) for 1 h, and then with primary antibodies including Ap2a (Santa Cruz, SC-8975); Chx10 (Bremner lab, University of Toronto); Cleaved caspase-3 (Cell Signaling 9661); Cone arrestin or ARR3 (Millipore, AB15282); γ-H2ax (Millipore, 05–636); Onecut2 or OC2 (R&D system AF6294); Ki67 (BD science Pharmingen 550609); PH3 (Upstate 05–598); Rhodopsin (Santa Cruz SC-57433) and Sox9 (Millipore AB5535), overnight at 4°C.

Techniques: Injection, Staining, Flow Cytometry, Control