rabbit antibodies against ing2  (Proteintech)

Journal: Frontiers in Cell and Developmental Biology

Article Title: ING2 Controls Mitochondrial Respiration via Modulating MRPL12 Ubiquitination in Renal Tubular Epithelial Cells

doi: 10.3389/fcell.2021.700195

Figure Lengend Snippet: ING2 positively regulates mitochondrial respiration in tubular epithelial cells. HK2 cells were transfected with ING2-shRNA or ING2-overexpression plasmid. (A,B) The transfection efficiencies were evaluated by qPCR and Western blotting, respectively. (C–F) Following transfection, mitochondrial OXPHOS of HK2 cells were detected using Seahorse. OCR, oxygen consumption rate; FCCP, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone; A&R, antimycin and rotenone. (G,H) mtDNA-encoded components for complex I, III, IV, and V and 16S rRNA were determined by qPCR. (I) mtDNA-encoded components for complex I, III, IV, and V were determined by Western blotting. (J) The mitochondria were stained with MitoTracker (red) and ING2 (green) followed by DAPI (blue) re-dyeing. (K) The mitochondria DNA (mtDNA) copy number was determined by qPCR with G6PC serving as internal reference. Data were from three individual experiments and presented as mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. ns, not significant.

Article Snippet: Sections were stained with rabbit antibodies against ING2 (1:200, Proteintech), MRPL12 (1:200, Proteintech), ND2 (1:200, Proteintech), or COX II (1:200, Proteintech) overnight at 4°C and incubated with the secondary antibody (HRP labeled goat anti-rabbit IgG, 1:1,000) for 1 h at RT.

Techniques: Transfection, shRNA, Over Expression, Plasmid Preparation, Western Blot, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: ING2 Controls Mitochondrial Respiration via Modulating MRPL12 Ubiquitination in Renal Tubular Epithelial Cells

doi: 10.3389/fcell.2021.700195

Figure Lengend Snippet: MRPL12 mediated the effects of ING2 on mitochondrial OXPHOS in tubular epithelial cells. (A,B) HK2 cells were transfected with ING2-shRNA or ING2-overexpression plasmid. The mRNA and protein levels of TFAM and MRPL12 were evaluated by qPCR and Western blotting. (C,D) HK2 cells were divided into control group, ING2 overexpression group, MRPL12 knockout group, and MRPL12 knockout plus ING2 overexpression group. Mitochondrial OXPHOS was detected using Seahorse. OCR, oxygen consumption rate; FCCP, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone; A&R, antimycin and rotenone. (E) Western blotting to validate the altered expression of mtDNA-encoded components of mitochondria complex. Data were from three individual experiments and presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001. ns, not significant.

Article Snippet: Sections were stained with rabbit antibodies against ING2 (1:200, Proteintech), MRPL12 (1:200, Proteintech), ND2 (1:200, Proteintech), or COX II (1:200, Proteintech) overnight at 4°C and incubated with the secondary antibody (HRP labeled goat anti-rabbit IgG, 1:1,000) for 1 h at RT.

Techniques: Transfection, shRNA, Over Expression, Plasmid Preparation, Western Blot, Knock-Out, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: ING2 Controls Mitochondrial Respiration via Modulating MRPL12 Ubiquitination in Renal Tubular Epithelial Cells

doi: 10.3389/fcell.2021.700195

Figure Lengend Snippet: ING2 inhibited the ubiquitination of MRPL12. (A) HK2 cells were transfected with control or ING2-shRNA, followed by MG132 treatment for 6 h. Protein levels of ING2 and MRPL12 were evaluated by Western blotting. (B) A ubiquitin modified site was found at 58 lysine residue (red, upper panel), predicted using predictor of protein ubiquitination sites, UbPred. (C,D) The ubiquitination of MRPL12 was determined by proximity ligation assay (PLA) and immunoprecipitation, respectively, after transfected ING2-overexpression plasmid. (E) By using ASEB, a web tool for predicting protein acetylation site, a acetylation site was detected at the 185 lysine residue (green, lower panel). (F) The acetylation of MRPL12 was determined by immunoprecipitation after transfected ING2-overexpression plasmid. IP, immunoprecipitation; WB, Western blotting. Data were from three individual experiments.

Article Snippet: Sections were stained with rabbit antibodies against ING2 (1:200, Proteintech), MRPL12 (1:200, Proteintech), ND2 (1:200, Proteintech), or COX II (1:200, Proteintech) overnight at 4°C and incubated with the secondary antibody (HRP labeled goat anti-rabbit IgG, 1:1,000) for 1 h at RT.

Techniques: Transfection, shRNA, Western Blot, Modification, Proximity Ligation Assay, Immunoprecipitation, Over Expression, Plasmid Preparation

Journal: Frontiers in Cell and Developmental Biology

Article Title: ING2 Controls Mitochondrial Respiration via Modulating MRPL12 Ubiquitination in Renal Tubular Epithelial Cells

doi: 10.3389/fcell.2021.700195

Figure Lengend Snippet: ING2 ameliorated the ischemia induced mitochondrial OXPHOS defects and tubular cell apoptosis. (A) Immunostaining of kidney tissue section from healthy individual and AKI patients with antibodies against ING2. At least two patients for each. (B) Immunostaining of kidney tissue section from healthy and AKI mice with antibodies against ING2 and MRPL12. HK2 cells were subjected to serum deprivation for 48 h, and protein contents of both ING2, MRPL12, and mtDNA-encoded components of mitochondria complex (C) were figured out by Western blotting. The apoptosis of HK2 cells was evaluated by flow cytometry (D) . HK2 cells were divided into control group, ING2 overexpression group, serum deprivation group, and serum deprivation plus ING2 overexpression group. MRPL12 and mtDNA-encoded components of mitochondria complex were detected through Western blotting (E) . Mitochondrial OXPHOS was detected using Seahorse (F,G) . The apoptosis of HK2 cells was evaluated by flow cytometry (H) , and the ubiquitination of MRPL12 was determined by proximity ligation assay (PLA) (I) . OCR, oxygen consumption rate; FCCP, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone; A&R, antimycin and rotenone. Data were from three individual experiments and presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001. ns, not significant.

Article Snippet: Sections were stained with rabbit antibodies against ING2 (1:200, Proteintech), MRPL12 (1:200, Proteintech), ND2 (1:200, Proteintech), or COX II (1:200, Proteintech) overnight at 4°C and incubated with the secondary antibody (HRP labeled goat anti-rabbit IgG, 1:1,000) for 1 h at RT.

Techniques: Immunostaining, Western Blot, Flow Cytometry, Over Expression, Proximity Ligation Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: ING2 Controls Mitochondrial Respiration via Modulating MRPL12 Ubiquitination in Renal Tubular Epithelial Cells

doi: 10.3389/fcell.2021.700195

Figure Lengend Snippet: ING2 overexpression effectively ameliorated ischemic kidney injury. The mice were injected with rAAV-con or rAAV-ING2 followed by the induction of IRI with sham operation group as control. (A) Immunofluorescent staining of kidneys after the intra-renal injection of rAAV vectors with ING2 (red) and DAPI (blue). (B) Immunostaining of renal tissue sections with antibodies against ING2, MRPL12, ND2, and COX II. (C) Western blot analysis of renal tissue ING2, MRPL12, ND2, and COX II. (D) Levels of creatinine in the serum or urine of IRI and sham operation mice. Data were from two individual experiments and presented as mean ± SEM. rAAV-con + IRI, n = 6; rAAV-ING2 + IRI, n = 6; rAAV-con + sham, n = 4; rAAV-ING2 + sham, n = 4. ** p < 0.01. ns, not significant.

Article Snippet: Sections were stained with rabbit antibodies against ING2 (1:200, Proteintech), MRPL12 (1:200, Proteintech), ND2 (1:200, Proteintech), or COX II (1:200, Proteintech) overnight at 4°C and incubated with the secondary antibody (HRP labeled goat anti-rabbit IgG, 1:1,000) for 1 h at RT.

Techniques: Over Expression, Injection, Staining, Immunostaining, Western Blot

Journal: The Indian Journal of Medical Research

Article Title: Downregulated inhibitor of growth 3 (ING3) expression during colorectal carcinogenesis

doi:

Figure Lengend Snippet: Immunostaining examination of ING3 protein in colorectal lesions. ING3 immunoreactivity was detectable in colorectal mucosa (A, B) , adenoma (C, D) , carcinoma (E, F) and infiltrating inflammatory cells (A, F) . (X 200 magnification).

Article Snippet: The rabbit anti-ING3 (Protein Tech Group Inc, USA; 1 : 100) was employed as the primary antibody.

Techniques: Immunostaining

Journal: The Indian Journal of Medical Research

Article Title: Downregulated inhibitor of growth 3 (ING3) expression during colorectal carcinogenesis

doi:

Figure Lengend Snippet: ING3 expression in colorectal non-neoplastic, adenomas and carcinoma

Article Snippet: The rabbit anti-ING3 (Protein Tech Group Inc, USA; 1 : 100) was employed as the primary antibody.

Techniques: Expressing

Journal: The Indian Journal of Medical Research

Article Title: Downregulated inhibitor of growth 3 (ING3) expression during colorectal carcinogenesis

doi:

Figure Lengend Snippet: Correlation between ING3 status and prognosis of the colorectal carcinoma patients. Kaplan-Meier curves for cumulative survival rate of patients with colorectal carcinomas according to ING3 expression status.

Article Snippet: The rabbit anti-ING3 (Protein Tech Group Inc, USA; 1 : 100) was employed as the primary antibody.

Techniques: Expressing

Journal: The Indian Journal of Medical Research

Article Title: Downregulated inhibitor of growth 3 (ING3) expression during colorectal carcinogenesis

doi:

Figure Lengend Snippet: Immunostaining examination of ING3 protein in colorectal lesions. ING3 immunoreactivity was detectable in colorectal mucosa (A, B) , adenoma (C, D) , carcinoma (E, F) and infiltrating inflammatory cells (A, F) . (X 200 magnification).

Article Snippet: For immunobloting, the membrane was incubated for 1 h with rabbit anti- ING3 antibody (Protein Tech Group Inc, USA; 1: 500) or mouse anti-β-actin (Santa Cruz, USA).

Techniques: Immunostaining

Journal: The Indian Journal of Medical Research

Article Title: Downregulated inhibitor of growth 3 (ING3) expression during colorectal carcinogenesis

doi:

Figure Lengend Snippet: ING3 expression in colorectal non-neoplastic, adenomas and carcinoma

Article Snippet: For immunobloting, the membrane was incubated for 1 h with rabbit anti- ING3 antibody (Protein Tech Group Inc, USA; 1: 500) or mouse anti-β-actin (Santa Cruz, USA).

Techniques: Expressing

Journal: The Indian Journal of Medical Research

Article Title: Downregulated inhibitor of growth 3 (ING3) expression during colorectal carcinogenesis

doi:

Figure Lengend Snippet: Correlation between ING3 status and prognosis of the colorectal carcinoma patients. Kaplan-Meier curves for cumulative survival rate of patients with colorectal carcinomas according to ING3 expression status.

Article Snippet: For immunobloting, the membrane was incubated for 1 h with rabbit anti- ING3 antibody (Protein Tech Group Inc, USA; 1: 500) or mouse anti-β-actin (Santa Cruz, USA).

Techniques: Expressing

Journal: Oncology Letters

Article Title: Downregulation of inhibitor of growth 3 is correlated with tumorigenesis and progression of hepatocellular carcinoma

doi: 10.3892/ol.2012.685

Figure Lengend Snippet: Expression pattern of ING3 in HCC samples and cell lines. (A) The transcript level of ING3 was measured in 49 paired HCC and adjacent non-cancerous livers by quantitative RT-PCR, where GAPDH was used as an internal reference. The line within each box is the median -ΔCt value; the upper and lower edges of each box show the 75th and 25th percentile, respectively. The upper and lower bars are the highest and lowest values. P-value was calculated by paired sample t-test (p<0.001). (B) Representative semi-quantitative RT-PCR of ING3 of 3 pairs of HCC samples. C, cancer tissue; N, adjacent noncancerous liver. (C) Relative mRNA level of ING3 was evaluated in HCC cell lines and fetal liver by semiquantitative RT-PCR. HCC, hepatocellular carcinoma; ING3, inhibitor of growth 3; RT-PCR, reverse transcription-polymerase chain reaction.

Article Snippet: The slides were incubated overnight at 4°C with rabbit anti-ING3 polyclonal antibody (1:200 dilution), followed by incubation with a horseradish peroxidase-conjugated anti-rabbit secondary antibody (Dako Japan Ltd., Kyoto, Japan) at 37°C for 30 min. Normal rabbit IgG was used as a negative control.

Techniques: Expressing, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: Downregulation of inhibitor of growth 3 is correlated with tumorigenesis and progression of hepatocellular carcinoma

doi: 10.3892/ol.2012.685

Figure Lengend Snippet: Representative immunohistochemical detection of the ING3 protein expression in HCC and surrounding noncancerous tissues. HCC, hepatocellular carcinoma; ING3, inhibitor of growth 3.

Article Snippet: The slides were incubated overnight at 4°C with rabbit anti-ING3 polyclonal antibody (1:200 dilution), followed by incubation with a horseradish peroxidase-conjugated anti-rabbit secondary antibody (Dako Japan Ltd., Kyoto, Japan) at 37°C for 30 min. Normal rabbit IgG was used as a negative control.

Techniques: Immunohistochemical staining, Expressing

Journal: Oncology Letters

Article Title: Downregulation of inhibitor of growth 3 is correlated with tumorigenesis and progression of hepatocellular carcinoma

doi: 10.3892/ol.2012.685

Figure Lengend Snippet: Correlations between ING3 expression and clinicopathological variables of 112 cases of HCC.

Article Snippet: The slides were incubated overnight at 4°C with rabbit anti-ING3 polyclonal antibody (1:200 dilution), followed by incubation with a horseradish peroxidase-conjugated anti-rabbit secondary antibody (Dako Japan Ltd., Kyoto, Japan) at 37°C for 30 min. Normal rabbit IgG was used as a negative control.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Downregulation of inhibitor of growth 3 is correlated with tumorigenesis and progression of hepatocellular carcinoma

doi: 10.3892/ol.2012.685

Figure Lengend Snippet: ING3 overexpression inhibits cell proliferation and colony formation. (A) Exogenous ING3 was expressed in Hep3B and Huh7 cells by transient transfection with pcDNA3.0-ING3, confirmed by western blot analysis. Cells transfected with an empty vector were used as the control. (B) The growth curve of Hep3B cells with the exogenous ING3. Cells transfected with the empty vector pcDNA3.0 served as controls. The experiments were repeated three times and the spots represent the average values of triplicate wells, with SD included for each mean value. (C) ING3 suppressed the ability of Hep3B cells to form colonies. The relative numbers of colonies of Hep3B cells transfected with pcDNA3.0-ING3 and a control vector were calculated from three independent experiments (***p<0.01). The lower panel showed the representative dishes of the experiments. (D) ING3 inhibited the colony formation of Huh7 cells (*p<0.05 as compared with the control vectors). ING3, inhibitor of growth 3.

Article Snippet: The slides were incubated overnight at 4°C with rabbit anti-ING3 polyclonal antibody (1:200 dilution), followed by incubation with a horseradish peroxidase-conjugated anti-rabbit secondary antibody (Dako Japan Ltd., Kyoto, Japan) at 37°C for 30 min. Normal rabbit IgG was used as a negative control.

Techniques: Over Expression, Transfection, Western Blot, Plasmid Preparation

Journal: Oncology Letters

Article Title: Downregulation of inhibitor of growth 3 is correlated with tumorigenesis and progression of hepatocellular carcinoma

doi: 10.3892/ol.2012.685

Figure Lengend Snippet: ING3 inhibits the migration of hepatoma cells. (A and B) ING3 overexpression reduced the migration of (A) Hep3B and (B) Huh7 cells. The two cell types were scraped with micropipette tips following transfection with pcDNA3.0-ING3 or pcDNA3.0. Images were captured at 0, 24 and 48 h following wounding. Cells transfected with an empty vector were used as controls. (C and D) The wound healing rates at 24 and 48 h following wounding of (C) Hep3B and (D) Huh7 cells. Values are the mean ± SD. *p<0.05. ING3, inhibitor of growth 3.

Article Snippet: The slides were incubated overnight at 4°C with rabbit anti-ING3 polyclonal antibody (1:200 dilution), followed by incubation with a horseradish peroxidase-conjugated anti-rabbit secondary antibody (Dako Japan Ltd., Kyoto, Japan) at 37°C for 30 min. Normal rabbit IgG was used as a negative control.

Techniques: Migration, Over Expression, Transfection, Plasmid Preparation

Journal: Oncology Letters

Article Title: Downregulation of inhibitor of growth 3 is correlated with tumorigenesis and progression of hepatocellular carcinoma

doi: 10.3892/ol.2012.685

Figure Lengend Snippet: Expression pattern of ING3 in HCC samples and cell lines. (A) The transcript level of ING3 was measured in 49 paired HCC and adjacent non-cancerous livers by quantitative RT-PCR, where GAPDH was used as an internal reference. The line within each box is the median -ΔCt value; the upper and lower edges of each box show the 75th and 25th percentile, respectively. The upper and lower bars are the highest and lowest values. P-value was calculated by paired sample t-test (p<0.001). (B) Representative semi-quantitative RT-PCR of ING3 of 3 pairs of HCC samples. C, cancer tissue; N, adjacent noncancerous liver. (C) Relative mRNA level of ING3 was evaluated in HCC cell lines and fetal liver by semiquantitative RT-PCR. HCC, hepatocellular carcinoma; ING3, inhibitor of growth 3; RT-PCR, reverse transcription-polymerase chain reaction.

Article Snippet: Rabbit anti-ING3 and mouse anti-β-actin (Sigma) antibodies were used in this study.

Techniques: Expressing, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: Downregulation of inhibitor of growth 3 is correlated with tumorigenesis and progression of hepatocellular carcinoma

doi: 10.3892/ol.2012.685

Figure Lengend Snippet: Representative immunohistochemical detection of the ING3 protein expression in HCC and surrounding noncancerous tissues. HCC, hepatocellular carcinoma; ING3, inhibitor of growth 3.

Article Snippet: Rabbit anti-ING3 and mouse anti-β-actin (Sigma) antibodies were used in this study.

Techniques: Immunohistochemical staining, Expressing

Journal: Oncology Letters

Article Title: Downregulation of inhibitor of growth 3 is correlated with tumorigenesis and progression of hepatocellular carcinoma

doi: 10.3892/ol.2012.685

Figure Lengend Snippet: Correlations between ING3 expression and clinicopathological variables of 112 cases of HCC.

Article Snippet: Rabbit anti-ING3 and mouse anti-β-actin (Sigma) antibodies were used in this study.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Downregulation of inhibitor of growth 3 is correlated with tumorigenesis and progression of hepatocellular carcinoma

doi: 10.3892/ol.2012.685

Figure Lengend Snippet: ING3 overexpression inhibits cell proliferation and colony formation. (A) Exogenous ING3 was expressed in Hep3B and Huh7 cells by transient transfection with pcDNA3.0-ING3, confirmed by western blot analysis. Cells transfected with an empty vector were used as the control. (B) The growth curve of Hep3B cells with the exogenous ING3. Cells transfected with the empty vector pcDNA3.0 served as controls. The experiments were repeated three times and the spots represent the average values of triplicate wells, with SD included for each mean value. (C) ING3 suppressed the ability of Hep3B cells to form colonies. The relative numbers of colonies of Hep3B cells transfected with pcDNA3.0-ING3 and a control vector were calculated from three independent experiments (***p<0.01). The lower panel showed the representative dishes of the experiments. (D) ING3 inhibited the colony formation of Huh7 cells (*p<0.05 as compared with the control vectors). ING3, inhibitor of growth 3.

Article Snippet: Rabbit anti-ING3 and mouse anti-β-actin (Sigma) antibodies were used in this study.

Techniques: Over Expression, Transfection, Western Blot, Plasmid Preparation

Journal: Oncology Letters

Article Title: Downregulation of inhibitor of growth 3 is correlated with tumorigenesis and progression of hepatocellular carcinoma

doi: 10.3892/ol.2012.685

Figure Lengend Snippet: ING3 inhibits the migration of hepatoma cells. (A and B) ING3 overexpression reduced the migration of (A) Hep3B and (B) Huh7 cells. The two cell types were scraped with micropipette tips following transfection with pcDNA3.0-ING3 or pcDNA3.0. Images were captured at 0, 24 and 48 h following wounding. Cells transfected with an empty vector were used as controls. (C and D) The wound healing rates at 24 and 48 h following wounding of (C) Hep3B and (D) Huh7 cells. Values are the mean ± SD. *p<0.05. ING3, inhibitor of growth 3.

Article Snippet: Rabbit anti-ING3 and mouse anti-β-actin (Sigma) antibodies were used in this study.

Techniques: Migration, Over Expression, Transfection, Plasmid Preparation