Structured Review

Proteintech phb2
Phb2, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/phb2/product/Proteintech
Average 86 stars, based on 1 article reviews
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phb2 - by Bioz Stars, 2024-07
86/100 stars

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Structured Review

Proteintech phb2
Effect of MPP + on <t>PHB2</t> protein expression and mitochondrial function in SH-SY5Y cells. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp, and then induced with 1 mM MPP + for 24 hours. (A, B) PHB2 protein concentration in SH-SY5Y cells after 1 mM MPP + treatment for 6, 12, 24, and 48 hours. (A) Western blots showing PHB2 protein expression in MPP + -treated cells at different times. (B) Quantitative analysis of PHB2 protein expression. (C–G) Protein expression changes of Nrf2, HO-1, NQO-1, and PHB2. (C) Western blots showing Nrf2, HO-1, NQO-1, and PHB2 protein expression in MPP + -treated and PHB2-shRNA- or PHB2-Over Exp-treated cells. (D–G) Quantitative analysis of Nrf2, HO-1, NQO-1, and PHB2 protein expression. (H, I) The effect of PHB2 expression on mitochondrial membrane potential induced by MPP + . (H) JC-1 staining of SH-SY5Y cells treated with PHB2-shRNA and PHB2-Over Exp. Original magnification 10×, scale bar: 50 μm. (I) Quantitative analysis of JC-1 staining: the green/red ratio reflects changes in the mitochondrial membrane potential. (J, K) The effect of changes in PHB2 expression on ROS production under MPP + induction. (J) Intracellular ROS fluorescence. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp and labeled with DCFH-DA (green fluorescence). Original magnification 4×, scale bar: 100 μm. (K) Quantitative analysis of ROS (DCFH-DA) in SH-SY5Y cells. (L) Electron microscopy images of morphological changes in mitochondria in PHB2-shRNA-treated and MPP + -induced cells. Red boxes show magnified mitochondria. Scale bars: 1 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . control group; # P < 0.05,## P < 0.01, vs . MPP + group (one-way analysis of variance with Tukey's multiple comparison test). DCFH-DA: 2′,7′-Dichlorodihydrofluorescein diacetate; HO-1: heme oxygenase-1; JC-1: 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide; MPP + : 1-methyl-4-phenylpyridinium; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; Over Exp: overexpression; PHB2: prohibitin 2; ROS: reactive oxygen species; SH-SY5Y: human neuroblastoma cell line.
Phb2, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/phb2/product/Proteintech
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
phb2 - by Bioz Stars, 2024-07
86/100 stars

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1) Product Images from "A novel mechanism of PHB2-mediated mitophagy participating in the development of Parkinson's disease"

Article Title: A novel mechanism of PHB2-mediated mitophagy participating in the development of Parkinson's disease

Journal: Neural Regeneration Research

doi: 10.4103/1673-5374.389356

Effect of MPP + on PHB2 protein expression and mitochondrial function in SH-SY5Y cells. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp, and then induced with 1 mM MPP + for 24 hours. (A, B) PHB2 protein concentration in SH-SY5Y cells after 1 mM MPP + treatment for 6, 12, 24, and 48 hours. (A) Western blots showing PHB2 protein expression in MPP + -treated cells at different times. (B) Quantitative analysis of PHB2 protein expression. (C–G) Protein expression changes of Nrf2, HO-1, NQO-1, and PHB2. (C) Western blots showing Nrf2, HO-1, NQO-1, and PHB2 protein expression in MPP + -treated and PHB2-shRNA- or PHB2-Over Exp-treated cells. (D–G) Quantitative analysis of Nrf2, HO-1, NQO-1, and PHB2 protein expression. (H, I) The effect of PHB2 expression on mitochondrial membrane potential induced by MPP + . (H) JC-1 staining of SH-SY5Y cells treated with PHB2-shRNA and PHB2-Over Exp. Original magnification 10×, scale bar: 50 μm. (I) Quantitative analysis of JC-1 staining: the green/red ratio reflects changes in the mitochondrial membrane potential. (J, K) The effect of changes in PHB2 expression on ROS production under MPP + induction. (J) Intracellular ROS fluorescence. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp and labeled with DCFH-DA (green fluorescence). Original magnification 4×, scale bar: 100 μm. (K) Quantitative analysis of ROS (DCFH-DA) in SH-SY5Y cells. (L) Electron microscopy images of morphological changes in mitochondria in PHB2-shRNA-treated and MPP + -induced cells. Red boxes show magnified mitochondria. Scale bars: 1 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . control group; # P < 0.05,## P < 0.01, vs . MPP + group (one-way analysis of variance with Tukey's multiple comparison test). DCFH-DA: 2′,7′-Dichlorodihydrofluorescein diacetate; HO-1: heme oxygenase-1; JC-1: 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide; MPP + : 1-methyl-4-phenylpyridinium; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; Over Exp: overexpression; PHB2: prohibitin 2; ROS: reactive oxygen species; SH-SY5Y: human neuroblastoma cell line.
Figure Legend Snippet: Effect of MPP + on PHB2 protein expression and mitochondrial function in SH-SY5Y cells. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp, and then induced with 1 mM MPP + for 24 hours. (A, B) PHB2 protein concentration in SH-SY5Y cells after 1 mM MPP + treatment for 6, 12, 24, and 48 hours. (A) Western blots showing PHB2 protein expression in MPP + -treated cells at different times. (B) Quantitative analysis of PHB2 protein expression. (C–G) Protein expression changes of Nrf2, HO-1, NQO-1, and PHB2. (C) Western blots showing Nrf2, HO-1, NQO-1, and PHB2 protein expression in MPP + -treated and PHB2-shRNA- or PHB2-Over Exp-treated cells. (D–G) Quantitative analysis of Nrf2, HO-1, NQO-1, and PHB2 protein expression. (H, I) The effect of PHB2 expression on mitochondrial membrane potential induced by MPP + . (H) JC-1 staining of SH-SY5Y cells treated with PHB2-shRNA and PHB2-Over Exp. Original magnification 10×, scale bar: 50 μm. (I) Quantitative analysis of JC-1 staining: the green/red ratio reflects changes in the mitochondrial membrane potential. (J, K) The effect of changes in PHB2 expression on ROS production under MPP + induction. (J) Intracellular ROS fluorescence. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp and labeled with DCFH-DA (green fluorescence). Original magnification 4×, scale bar: 100 μm. (K) Quantitative analysis of ROS (DCFH-DA) in SH-SY5Y cells. (L) Electron microscopy images of morphological changes in mitochondria in PHB2-shRNA-treated and MPP + -induced cells. Red boxes show magnified mitochondria. Scale bars: 1 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . control group; # P < 0.05,## P < 0.01, vs . MPP + group (one-way analysis of variance with Tukey's multiple comparison test). DCFH-DA: 2′,7′-Dichlorodihydrofluorescein diacetate; HO-1: heme oxygenase-1; JC-1: 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide; MPP + : 1-methyl-4-phenylpyridinium; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; Over Exp: overexpression; PHB2: prohibitin 2; ROS: reactive oxygen species; SH-SY5Y: human neuroblastoma cell line.

Techniques Used: Expressing, shRNA, Protein Concentration, Western Blot, Membrane, Staining, Fluorescence, Labeling, Electron Microscopy, Comparison, Over Expression

Effect of PHB2 on mitophagy in a PD cell model. (A–F) PHB2 and LC3II/LC3I protein expression in SH-SY5Y cells after treatment with PHB2-shRNA or PHB2-Over Exp and MPP + (1 mM, 24 hours). (A) Protein expression levels of PHB2 and LC3 in SH-SY5Y cells treated with PHB2-shRNA under MPP + induction. (B, C) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (D) Protein expression levels of PHB2 and LC3 in SH-SY5Y cells treated with PHB2-Over Exp under MPP + induction. (E, F) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (G) Cellular localization of LC3 and TOM20 or TIM23 in SH-SY5Y cells treated with PHB2-shRNA under MPP + induction. Immunofluorescence changes of LC3 and mitochondrial proteins (TOM20 and TIM23). Red fluorescence represents TOM20 and TIM23, green fluorescence represents LC3, and blue fluorescence represents DAPI. Original magnification 20×, Scale bars: 20 μm. Data are expressed as mean ± SEM ( n = 3). ** P < 0.01, *** P < 0.001, vs . control group (one-way analysis of variance with Tukey's multiple comparison test). DAPI: 4′,6-Diamidino-2-phenylindole; LC3: microtubule-associated protein 1 light chain 3; MPP + : 1-methyl-4-phenylpyridinium; Over Exp: overexpression; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line; TIM23: translocase of inner mitochondrial membrane 23; TOM20: translocase of outer mitochondrial membrane 20.
Figure Legend Snippet: Effect of PHB2 on mitophagy in a PD cell model. (A–F) PHB2 and LC3II/LC3I protein expression in SH-SY5Y cells after treatment with PHB2-shRNA or PHB2-Over Exp and MPP + (1 mM, 24 hours). (A) Protein expression levels of PHB2 and LC3 in SH-SY5Y cells treated with PHB2-shRNA under MPP + induction. (B, C) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (D) Protein expression levels of PHB2 and LC3 in SH-SY5Y cells treated with PHB2-Over Exp under MPP + induction. (E, F) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (G) Cellular localization of LC3 and TOM20 or TIM23 in SH-SY5Y cells treated with PHB2-shRNA under MPP + induction. Immunofluorescence changes of LC3 and mitochondrial proteins (TOM20 and TIM23). Red fluorescence represents TOM20 and TIM23, green fluorescence represents LC3, and blue fluorescence represents DAPI. Original magnification 20×, Scale bars: 20 μm. Data are expressed as mean ± SEM ( n = 3). ** P < 0.01, *** P < 0.001, vs . control group (one-way analysis of variance with Tukey's multiple comparison test). DAPI: 4′,6-Diamidino-2-phenylindole; LC3: microtubule-associated protein 1 light chain 3; MPP + : 1-methyl-4-phenylpyridinium; Over Exp: overexpression; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line; TIM23: translocase of inner mitochondrial membrane 23; TOM20: translocase of outer mitochondrial membrane 20.

Techniques Used: Expressing, shRNA, Immunofluorescence, Fluorescence, Comparison, Over Expression, Membrane

Parkin increases PHB2/LC3-mediated mitophagy in SH-SY5Y cells. (A–D) Protein levels of Parkin, PHB2, and autophagy marker (LC3) in SH-SY5Y cells after treatment with Parkin-shRNA, Parkin-Over Exp, and MPP + (1 mM, 24 hours). (A) Protein expression levels of Parkin, LC3, and PHB2 in SH-SY5Y cells treated with Parkin-shRNA or Parkin-Over Exp under MPP + induction. (B–D) Quantitative analysis of Parkin, and LC3II/LC3I, PHB2 protein expression. (E) Immunofluorescence analysis of PHB2 and LC3. Cellular localization of LC3 and PHB2 in SH-SY5Y cells treated with Parkin-shRNA or Parkin-Over Exp under MPP + induction. Red fluorescence represents PHB2, green fluorescence represents LC3, and blue fluorescence represents DAPI. Original magnification 40×, Scale bar: 20 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs. control group; # P < 0.05,### P < 0.001, vs . MPP + group (one-way analysis of variance with Tukey's multiple comparison test). DAPI: 4′,6-Diamidino-2-phenylindole; LC3: microtubule-associated protein 1 light chain 3; MPP + : 1-methyl-4-phenylpyridinium; Over Exp: overexpression; Parkin: parkin RBR E3 ubiquitin-protein ligase; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line.
Figure Legend Snippet: Parkin increases PHB2/LC3-mediated mitophagy in SH-SY5Y cells. (A–D) Protein levels of Parkin, PHB2, and autophagy marker (LC3) in SH-SY5Y cells after treatment with Parkin-shRNA, Parkin-Over Exp, and MPP + (1 mM, 24 hours). (A) Protein expression levels of Parkin, LC3, and PHB2 in SH-SY5Y cells treated with Parkin-shRNA or Parkin-Over Exp under MPP + induction. (B–D) Quantitative analysis of Parkin, and LC3II/LC3I, PHB2 protein expression. (E) Immunofluorescence analysis of PHB2 and LC3. Cellular localization of LC3 and PHB2 in SH-SY5Y cells treated with Parkin-shRNA or Parkin-Over Exp under MPP + induction. Red fluorescence represents PHB2, green fluorescence represents LC3, and blue fluorescence represents DAPI. Original magnification 40×, Scale bar: 20 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs. control group; # P < 0.05,### P < 0.001, vs . MPP + group (one-way analysis of variance with Tukey's multiple comparison test). DAPI: 4′,6-Diamidino-2-phenylindole; LC3: microtubule-associated protein 1 light chain 3; MPP + : 1-methyl-4-phenylpyridinium; Over Exp: overexpression; Parkin: parkin RBR E3 ubiquitin-protein ligase; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line.

Techniques Used: Marker, shRNA, Expressing, Immunofluorescence, Fluorescence, Comparison, Over Expression

Parkin regulates anti-oxidative stress protein expression via PHB2. (A–D) SH-SY5Y cells were treated with PHB2-shRNA, Parkin-Over Exp, and MPP + (1 mM, 24 hours). Changes in Nrf2, HO-1, and NQO-1 protein levels and quantitative analysis. (A) Protein expression levels of Nrf2, HO-1, and NQO-1 in SH-SY5Y cells treated with PHB2-shRNA or Parkin-Over Exp under MPP + induction. (B–D) Quantitative analysis of Nrf2, HO-1, and NQO-1 protein expression. Data are expressed as mean ± SEM ( n = 3). *** P < 0.001, vs. control group; # P < 0.05,### P < 0.001, vs. MPP + group (one-way analysis of variance with Tukey's multiple comparison test). HO-1: Heme oxygenase-1; MPP + : 1-methyl-4-phenylpyridinium; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; Over Exp: overexpression; Parkin: parkin RBR E3 ubiquitin-protein ligase; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line.
Figure Legend Snippet: Parkin regulates anti-oxidative stress protein expression via PHB2. (A–D) SH-SY5Y cells were treated with PHB2-shRNA, Parkin-Over Exp, and MPP + (1 mM, 24 hours). Changes in Nrf2, HO-1, and NQO-1 protein levels and quantitative analysis. (A) Protein expression levels of Nrf2, HO-1, and NQO-1 in SH-SY5Y cells treated with PHB2-shRNA or Parkin-Over Exp under MPP + induction. (B–D) Quantitative analysis of Nrf2, HO-1, and NQO-1 protein expression. Data are expressed as mean ± SEM ( n = 3). *** P < 0.001, vs. control group; # P < 0.05,### P < 0.001, vs. MPP + group (one-way analysis of variance with Tukey's multiple comparison test). HO-1: Heme oxygenase-1; MPP + : 1-methyl-4-phenylpyridinium; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; Over Exp: overexpression; Parkin: parkin RBR E3 ubiquitin-protein ligase; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line.

Techniques Used: Expressing, shRNA, Comparison, Over Expression

Silencing PHB2 inhibits mitophagy and aggravates dopaminergic neuronal loss in PD mice. (A) Schematic of PHB2-shRNA and MPTP treatment in mice. (B, C) Quantitative analysis of PHB2 protein levels in the midbrain of C57BL/6J mice after PHB2-shRNA injection. (B) Protein expression levels of PHB2 in C57BL/6J mice treated with PHB2-shRNA. (C) Quantitative analysis of PHB2 protein expression. (D–F) An acute PD model was established by intraperitoneal injection of MPTP in C57BL/6J mice after PHB2-shRNA injection. Changes and quantitative analysis of LC3II/LC3I and PHB2 protein levels in the midbrain. (D) Protein expression levels of PHB2 and LC3 after MPTP injection in PD model mice. (E, F) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (G, H) TH immunofluorescence (Alexa Fluor 488, green fluorescence) shows dopaminergic neurons in the substantia nigra in PD mice with silenced PHB2. (G) Immunofluorescence of TH-positive neurons in the substantia nigra. Mice were treated with PHB2-shRNA and MPTP. Original magnification 10×, Scale bar: 200 μm. (H) Quantitative analysis of TH-positive neurons in the substantia nigra. (I) Fluorescence co-localization of LC3 and PHB2, TIM23, or TOM20 in the substantia nigra. Mice were treated with PHB2-shRNA and MPTP. Red fluorescence (Alexa Fluor 555): PHB2, TOM20, and TIM23, green fluorescence (Alexa Fluor 488): LC3, and blue fluorescence: DAPI. Original magnification 20×, Scale bars: 20 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, *** P < 0.001, vs. control group; $ P < 0.05, $$ P < 0.01, $$$ P < 0.001, vs. MPTP group (one-way analysis of variance with Tukey's multiple comparison test). LC3: Microtubule-associated protein 1 light chain 3; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; PD: Parkinson's disease; PHB2: Prohibitin 2; TH: tyrosine hydroxylase; TIM23: translocase of inner mitochondrial membrane 23; TOM20: translocase of outer mitochondrial membrane 20.
Figure Legend Snippet: Silencing PHB2 inhibits mitophagy and aggravates dopaminergic neuronal loss in PD mice. (A) Schematic of PHB2-shRNA and MPTP treatment in mice. (B, C) Quantitative analysis of PHB2 protein levels in the midbrain of C57BL/6J mice after PHB2-shRNA injection. (B) Protein expression levels of PHB2 in C57BL/6J mice treated with PHB2-shRNA. (C) Quantitative analysis of PHB2 protein expression. (D–F) An acute PD model was established by intraperitoneal injection of MPTP in C57BL/6J mice after PHB2-shRNA injection. Changes and quantitative analysis of LC3II/LC3I and PHB2 protein levels in the midbrain. (D) Protein expression levels of PHB2 and LC3 after MPTP injection in PD model mice. (E, F) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (G, H) TH immunofluorescence (Alexa Fluor 488, green fluorescence) shows dopaminergic neurons in the substantia nigra in PD mice with silenced PHB2. (G) Immunofluorescence of TH-positive neurons in the substantia nigra. Mice were treated with PHB2-shRNA and MPTP. Original magnification 10×, Scale bar: 200 μm. (H) Quantitative analysis of TH-positive neurons in the substantia nigra. (I) Fluorescence co-localization of LC3 and PHB2, TIM23, or TOM20 in the substantia nigra. Mice were treated with PHB2-shRNA and MPTP. Red fluorescence (Alexa Fluor 555): PHB2, TOM20, and TIM23, green fluorescence (Alexa Fluor 488): LC3, and blue fluorescence: DAPI. Original magnification 20×, Scale bars: 20 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, *** P < 0.001, vs. control group; $ P < 0.05, $$ P < 0.01, $$$ P < 0.001, vs. MPTP group (one-way analysis of variance with Tukey's multiple comparison test). LC3: Microtubule-associated protein 1 light chain 3; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; PD: Parkinson's disease; PHB2: Prohibitin 2; TH: tyrosine hydroxylase; TIM23: translocase of inner mitochondrial membrane 23; TOM20: translocase of outer mitochondrial membrane 20.

Techniques Used: shRNA, Injection, Expressing, Immunofluorescence, Fluorescence, Comparison, Membrane

Silencing PHB2 reduces antioxidative stress protein expression and aggravates motor defecits in PD mice. (A–D) Changes and quantitative analysis of Nrf2, HO-1, and NQO-1 protein levels in the midbrain of PHB2-shRNA and MPTP-treated PD mice. (E) Quantitative analysis of mice in the tail suspension and rotarod tests. Data are expressed as mean ± SEM ( n = 5). * P < 0.05, ** P < 0.01, vs. control group; $ P < 0.05, $$$ P < 0.001, vs . MPTP group (one-way analysis of variance with Tukey's multiple comparison test). HO-1: Heme oxygenase-1; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; PD: Parkinson's disease; PHB2: prohibitin 2.
Figure Legend Snippet: Silencing PHB2 reduces antioxidative stress protein expression and aggravates motor defecits in PD mice. (A–D) Changes and quantitative analysis of Nrf2, HO-1, and NQO-1 protein levels in the midbrain of PHB2-shRNA and MPTP-treated PD mice. (E) Quantitative analysis of mice in the tail suspension and rotarod tests. Data are expressed as mean ± SEM ( n = 5). * P < 0.05, ** P < 0.01, vs. control group; $ P < 0.05, $$$ P < 0.001, vs . MPTP group (one-way analysis of variance with Tukey's multiple comparison test). HO-1: Heme oxygenase-1; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; PD: Parkinson's disease; PHB2: prohibitin 2.

Techniques Used: Expressing, shRNA, Suspension, Comparison


Structured Review

Proteintech antibodies against phb2
Effect of MPP + on <t>PHB2</t> protein expression and mitochondrial function in SH-SY5Y cells. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp, and then induced with 1 mM MPP + for 24 hours. (A, B) PHB2 protein concentration in SH-SY5Y cells after 1 mM MPP + treatment for 6, 12, 24, and 48 hours. (A) Western blots showing PHB2 protein expression in MPP + -treated cells at different times. (B) Quantitative analysis of PHB2 protein expression. (C–G) Protein expression changes of Nrf2, HO-1, NQO-1, and PHB2. (C) Western blots showing Nrf2, HO-1, NQO-1, and PHB2 protein expression in MPP + -treated and PHB2-shRNA- or PHB2-Over Exp-treated cells. (D–G) Quantitative analysis of Nrf2, HO-1, NQO-1, and PHB2 protein expression. (H, I) The effect of PHB2 expression on mitochondrial membrane potential induced by MPP + . (H) JC-1 staining of SH-SY5Y cells treated with PHB2-shRNA and PHB2-Over Exp. Original magnification 10×, scale bar: 50 μm. (I) Quantitative analysis of JC-1 staining: the green/red ratio reflects changes in the mitochondrial membrane potential. (J, K) The effect of changes in PHB2 expression on ROS production under MPP + induction. (J) Intracellular ROS fluorescence. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp and labeled with DCFH-DA (green fluorescence). Original magnification 4×, scale bar: 100 μm. (K) Quantitative analysis of ROS (DCFH-DA) in SH-SY5Y cells. (L) Electron microscopy images of morphological changes in mitochondria in PHB2-shRNA-treated and MPP + -induced cells. Red boxes show magnified mitochondria. Scale bars: 1 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . control group; # P < 0.05,## P < 0.01, vs . MPP + group (one-way analysis of variance with Tukey's multiple comparison test). DCFH-DA: 2′,7′-Dichlorodihydrofluorescein diacetate; HO-1: heme oxygenase-1; JC-1: 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide; MPP + : 1-methyl-4-phenylpyridinium; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; Over Exp: overexpression; PHB2: prohibitin 2; ROS: reactive oxygen species; SH-SY5Y: human neuroblastoma cell line.
Antibodies Against Phb2, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/antibodies against phb2/product/Proteintech
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
antibodies against phb2 - by Bioz Stars, 2024-07
86/100 stars

Images

1) Product Images from "A novel mechanism of PHB2-mediated mitophagy participating in the development of Parkinson's disease"

Article Title: A novel mechanism of PHB2-mediated mitophagy participating in the development of Parkinson's disease

Journal: Neural Regeneration Research

doi: 10.4103/1673-5374.389356

Effect of MPP + on PHB2 protein expression and mitochondrial function in SH-SY5Y cells. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp, and then induced with 1 mM MPP + for 24 hours. (A, B) PHB2 protein concentration in SH-SY5Y cells after 1 mM MPP + treatment for 6, 12, 24, and 48 hours. (A) Western blots showing PHB2 protein expression in MPP + -treated cells at different times. (B) Quantitative analysis of PHB2 protein expression. (C–G) Protein expression changes of Nrf2, HO-1, NQO-1, and PHB2. (C) Western blots showing Nrf2, HO-1, NQO-1, and PHB2 protein expression in MPP + -treated and PHB2-shRNA- or PHB2-Over Exp-treated cells. (D–G) Quantitative analysis of Nrf2, HO-1, NQO-1, and PHB2 protein expression. (H, I) The effect of PHB2 expression on mitochondrial membrane potential induced by MPP + . (H) JC-1 staining of SH-SY5Y cells treated with PHB2-shRNA and PHB2-Over Exp. Original magnification 10×, scale bar: 50 μm. (I) Quantitative analysis of JC-1 staining: the green/red ratio reflects changes in the mitochondrial membrane potential. (J, K) The effect of changes in PHB2 expression on ROS production under MPP + induction. (J) Intracellular ROS fluorescence. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp and labeled with DCFH-DA (green fluorescence). Original magnification 4×, scale bar: 100 μm. (K) Quantitative analysis of ROS (DCFH-DA) in SH-SY5Y cells. (L) Electron microscopy images of morphological changes in mitochondria in PHB2-shRNA-treated and MPP + -induced cells. Red boxes show magnified mitochondria. Scale bars: 1 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . control group; # P < 0.05,## P < 0.01, vs . MPP + group (one-way analysis of variance with Tukey's multiple comparison test). DCFH-DA: 2′,7′-Dichlorodihydrofluorescein diacetate; HO-1: heme oxygenase-1; JC-1: 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide; MPP + : 1-methyl-4-phenylpyridinium; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; Over Exp: overexpression; PHB2: prohibitin 2; ROS: reactive oxygen species; SH-SY5Y: human neuroblastoma cell line.
Figure Legend Snippet: Effect of MPP + on PHB2 protein expression and mitochondrial function in SH-SY5Y cells. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp, and then induced with 1 mM MPP + for 24 hours. (A, B) PHB2 protein concentration in SH-SY5Y cells after 1 mM MPP + treatment for 6, 12, 24, and 48 hours. (A) Western blots showing PHB2 protein expression in MPP + -treated cells at different times. (B) Quantitative analysis of PHB2 protein expression. (C–G) Protein expression changes of Nrf2, HO-1, NQO-1, and PHB2. (C) Western blots showing Nrf2, HO-1, NQO-1, and PHB2 protein expression in MPP + -treated and PHB2-shRNA- or PHB2-Over Exp-treated cells. (D–G) Quantitative analysis of Nrf2, HO-1, NQO-1, and PHB2 protein expression. (H, I) The effect of PHB2 expression on mitochondrial membrane potential induced by MPP + . (H) JC-1 staining of SH-SY5Y cells treated with PHB2-shRNA and PHB2-Over Exp. Original magnification 10×, scale bar: 50 μm. (I) Quantitative analysis of JC-1 staining: the green/red ratio reflects changes in the mitochondrial membrane potential. (J, K) The effect of changes in PHB2 expression on ROS production under MPP + induction. (J) Intracellular ROS fluorescence. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp and labeled with DCFH-DA (green fluorescence). Original magnification 4×, scale bar: 100 μm. (K) Quantitative analysis of ROS (DCFH-DA) in SH-SY5Y cells. (L) Electron microscopy images of morphological changes in mitochondria in PHB2-shRNA-treated and MPP + -induced cells. Red boxes show magnified mitochondria. Scale bars: 1 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . control group; # P < 0.05,## P < 0.01, vs . MPP + group (one-way analysis of variance with Tukey's multiple comparison test). DCFH-DA: 2′,7′-Dichlorodihydrofluorescein diacetate; HO-1: heme oxygenase-1; JC-1: 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide; MPP + : 1-methyl-4-phenylpyridinium; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; Over Exp: overexpression; PHB2: prohibitin 2; ROS: reactive oxygen species; SH-SY5Y: human neuroblastoma cell line.

Techniques Used: Expressing, shRNA, Protein Concentration, Western Blot, Membrane, Staining, Fluorescence, Labeling, Electron Microscopy, Comparison, Over Expression

Effect of PHB2 on mitophagy in a PD cell model. (A–F) PHB2 and LC3II/LC3I protein expression in SH-SY5Y cells after treatment with PHB2-shRNA or PHB2-Over Exp and MPP + (1 mM, 24 hours). (A) Protein expression levels of PHB2 and LC3 in SH-SY5Y cells treated with PHB2-shRNA under MPP + induction. (B, C) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (D) Protein expression levels of PHB2 and LC3 in SH-SY5Y cells treated with PHB2-Over Exp under MPP + induction. (E, F) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (G) Cellular localization of LC3 and TOM20 or TIM23 in SH-SY5Y cells treated with PHB2-shRNA under MPP + induction. Immunofluorescence changes of LC3 and mitochondrial proteins (TOM20 and TIM23). Red fluorescence represents TOM20 and TIM23, green fluorescence represents LC3, and blue fluorescence represents DAPI. Original magnification 20×, Scale bars: 20 μm. Data are expressed as mean ± SEM ( n = 3). ** P < 0.01, *** P < 0.001, vs . control group (one-way analysis of variance with Tukey's multiple comparison test). DAPI: 4′,6-Diamidino-2-phenylindole; LC3: microtubule-associated protein 1 light chain 3; MPP + : 1-methyl-4-phenylpyridinium; Over Exp: overexpression; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line; TIM23: translocase of inner mitochondrial membrane 23; TOM20: translocase of outer mitochondrial membrane 20.
Figure Legend Snippet: Effect of PHB2 on mitophagy in a PD cell model. (A–F) PHB2 and LC3II/LC3I protein expression in SH-SY5Y cells after treatment with PHB2-shRNA or PHB2-Over Exp and MPP + (1 mM, 24 hours). (A) Protein expression levels of PHB2 and LC3 in SH-SY5Y cells treated with PHB2-shRNA under MPP + induction. (B, C) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (D) Protein expression levels of PHB2 and LC3 in SH-SY5Y cells treated with PHB2-Over Exp under MPP + induction. (E, F) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (G) Cellular localization of LC3 and TOM20 or TIM23 in SH-SY5Y cells treated with PHB2-shRNA under MPP + induction. Immunofluorescence changes of LC3 and mitochondrial proteins (TOM20 and TIM23). Red fluorescence represents TOM20 and TIM23, green fluorescence represents LC3, and blue fluorescence represents DAPI. Original magnification 20×, Scale bars: 20 μm. Data are expressed as mean ± SEM ( n = 3). ** P < 0.01, *** P < 0.001, vs . control group (one-way analysis of variance with Tukey's multiple comparison test). DAPI: 4′,6-Diamidino-2-phenylindole; LC3: microtubule-associated protein 1 light chain 3; MPP + : 1-methyl-4-phenylpyridinium; Over Exp: overexpression; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line; TIM23: translocase of inner mitochondrial membrane 23; TOM20: translocase of outer mitochondrial membrane 20.

Techniques Used: Expressing, shRNA, Immunofluorescence, Fluorescence, Comparison, Over Expression, Membrane

Parkin increases PHB2/LC3-mediated mitophagy in SH-SY5Y cells. (A–D) Protein levels of Parkin, PHB2, and autophagy marker (LC3) in SH-SY5Y cells after treatment with Parkin-shRNA, Parkin-Over Exp, and MPP + (1 mM, 24 hours). (A) Protein expression levels of Parkin, LC3, and PHB2 in SH-SY5Y cells treated with Parkin-shRNA or Parkin-Over Exp under MPP + induction. (B–D) Quantitative analysis of Parkin, and LC3II/LC3I, PHB2 protein expression. (E) Immunofluorescence analysis of PHB2 and LC3. Cellular localization of LC3 and PHB2 in SH-SY5Y cells treated with Parkin-shRNA or Parkin-Over Exp under MPP + induction. Red fluorescence represents PHB2, green fluorescence represents LC3, and blue fluorescence represents DAPI. Original magnification 40×, Scale bar: 20 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs. control group; # P < 0.05,### P < 0.001, vs . MPP + group (one-way analysis of variance with Tukey's multiple comparison test). DAPI: 4′,6-Diamidino-2-phenylindole; LC3: microtubule-associated protein 1 light chain 3; MPP + : 1-methyl-4-phenylpyridinium; Over Exp: overexpression; Parkin: parkin RBR E3 ubiquitin-protein ligase; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line.
Figure Legend Snippet: Parkin increases PHB2/LC3-mediated mitophagy in SH-SY5Y cells. (A–D) Protein levels of Parkin, PHB2, and autophagy marker (LC3) in SH-SY5Y cells after treatment with Parkin-shRNA, Parkin-Over Exp, and MPP + (1 mM, 24 hours). (A) Protein expression levels of Parkin, LC3, and PHB2 in SH-SY5Y cells treated with Parkin-shRNA or Parkin-Over Exp under MPP + induction. (B–D) Quantitative analysis of Parkin, and LC3II/LC3I, PHB2 protein expression. (E) Immunofluorescence analysis of PHB2 and LC3. Cellular localization of LC3 and PHB2 in SH-SY5Y cells treated with Parkin-shRNA or Parkin-Over Exp under MPP + induction. Red fluorescence represents PHB2, green fluorescence represents LC3, and blue fluorescence represents DAPI. Original magnification 40×, Scale bar: 20 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs. control group; # P < 0.05,### P < 0.001, vs . MPP + group (one-way analysis of variance with Tukey's multiple comparison test). DAPI: 4′,6-Diamidino-2-phenylindole; LC3: microtubule-associated protein 1 light chain 3; MPP + : 1-methyl-4-phenylpyridinium; Over Exp: overexpression; Parkin: parkin RBR E3 ubiquitin-protein ligase; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line.

Techniques Used: Marker, shRNA, Expressing, Immunofluorescence, Fluorescence, Comparison, Over Expression

Parkin regulates anti-oxidative stress protein expression via PHB2. (A–D) SH-SY5Y cells were treated with PHB2-shRNA, Parkin-Over Exp, and MPP + (1 mM, 24 hours). Changes in Nrf2, HO-1, and NQO-1 protein levels and quantitative analysis. (A) Protein expression levels of Nrf2, HO-1, and NQO-1 in SH-SY5Y cells treated with PHB2-shRNA or Parkin-Over Exp under MPP + induction. (B–D) Quantitative analysis of Nrf2, HO-1, and NQO-1 protein expression. Data are expressed as mean ± SEM ( n = 3). *** P < 0.001, vs. control group; # P < 0.05,### P < 0.001, vs. MPP + group (one-way analysis of variance with Tukey's multiple comparison test). HO-1: Heme oxygenase-1; MPP + : 1-methyl-4-phenylpyridinium; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; Over Exp: overexpression; Parkin: parkin RBR E3 ubiquitin-protein ligase; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line.
Figure Legend Snippet: Parkin regulates anti-oxidative stress protein expression via PHB2. (A–D) SH-SY5Y cells were treated with PHB2-shRNA, Parkin-Over Exp, and MPP + (1 mM, 24 hours). Changes in Nrf2, HO-1, and NQO-1 protein levels and quantitative analysis. (A) Protein expression levels of Nrf2, HO-1, and NQO-1 in SH-SY5Y cells treated with PHB2-shRNA or Parkin-Over Exp under MPP + induction. (B–D) Quantitative analysis of Nrf2, HO-1, and NQO-1 protein expression. Data are expressed as mean ± SEM ( n = 3). *** P < 0.001, vs. control group; # P < 0.05,### P < 0.001, vs. MPP + group (one-way analysis of variance with Tukey's multiple comparison test). HO-1: Heme oxygenase-1; MPP + : 1-methyl-4-phenylpyridinium; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; Over Exp: overexpression; Parkin: parkin RBR E3 ubiquitin-protein ligase; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line.

Techniques Used: Expressing, shRNA, Comparison, Over Expression

Silencing PHB2 inhibits mitophagy and aggravates dopaminergic neuronal loss in PD mice. (A) Schematic of PHB2-shRNA and MPTP treatment in mice. (B, C) Quantitative analysis of PHB2 protein levels in the midbrain of C57BL/6J mice after PHB2-shRNA injection. (B) Protein expression levels of PHB2 in C57BL/6J mice treated with PHB2-shRNA. (C) Quantitative analysis of PHB2 protein expression. (D–F) An acute PD model was established by intraperitoneal injection of MPTP in C57BL/6J mice after PHB2-shRNA injection. Changes and quantitative analysis of LC3II/LC3I and PHB2 protein levels in the midbrain. (D) Protein expression levels of PHB2 and LC3 after MPTP injection in PD model mice. (E, F) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (G, H) TH immunofluorescence (Alexa Fluor 488, green fluorescence) shows dopaminergic neurons in the substantia nigra in PD mice with silenced PHB2. (G) Immunofluorescence of TH-positive neurons in the substantia nigra. Mice were treated with PHB2-shRNA and MPTP. Original magnification 10×, Scale bar: 200 μm. (H) Quantitative analysis of TH-positive neurons in the substantia nigra. (I) Fluorescence co-localization of LC3 and PHB2, TIM23, or TOM20 in the substantia nigra. Mice were treated with PHB2-shRNA and MPTP. Red fluorescence (Alexa Fluor 555): PHB2, TOM20, and TIM23, green fluorescence (Alexa Fluor 488): LC3, and blue fluorescence: DAPI. Original magnification 20×, Scale bars: 20 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, *** P < 0.001, vs. control group; $ P < 0.05, $$ P < 0.01, $$$ P < 0.001, vs. MPTP group (one-way analysis of variance with Tukey's multiple comparison test). LC3: Microtubule-associated protein 1 light chain 3; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; PD: Parkinson's disease; PHB2: Prohibitin 2; TH: tyrosine hydroxylase; TIM23: translocase of inner mitochondrial membrane 23; TOM20: translocase of outer mitochondrial membrane 20.
Figure Legend Snippet: Silencing PHB2 inhibits mitophagy and aggravates dopaminergic neuronal loss in PD mice. (A) Schematic of PHB2-shRNA and MPTP treatment in mice. (B, C) Quantitative analysis of PHB2 protein levels in the midbrain of C57BL/6J mice after PHB2-shRNA injection. (B) Protein expression levels of PHB2 in C57BL/6J mice treated with PHB2-shRNA. (C) Quantitative analysis of PHB2 protein expression. (D–F) An acute PD model was established by intraperitoneal injection of MPTP in C57BL/6J mice after PHB2-shRNA injection. Changes and quantitative analysis of LC3II/LC3I and PHB2 protein levels in the midbrain. (D) Protein expression levels of PHB2 and LC3 after MPTP injection in PD model mice. (E, F) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (G, H) TH immunofluorescence (Alexa Fluor 488, green fluorescence) shows dopaminergic neurons in the substantia nigra in PD mice with silenced PHB2. (G) Immunofluorescence of TH-positive neurons in the substantia nigra. Mice were treated with PHB2-shRNA and MPTP. Original magnification 10×, Scale bar: 200 μm. (H) Quantitative analysis of TH-positive neurons in the substantia nigra. (I) Fluorescence co-localization of LC3 and PHB2, TIM23, or TOM20 in the substantia nigra. Mice were treated with PHB2-shRNA and MPTP. Red fluorescence (Alexa Fluor 555): PHB2, TOM20, and TIM23, green fluorescence (Alexa Fluor 488): LC3, and blue fluorescence: DAPI. Original magnification 20×, Scale bars: 20 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, *** P < 0.001, vs. control group; $ P < 0.05, $$ P < 0.01, $$$ P < 0.001, vs. MPTP group (one-way analysis of variance with Tukey's multiple comparison test). LC3: Microtubule-associated protein 1 light chain 3; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; PD: Parkinson's disease; PHB2: Prohibitin 2; TH: tyrosine hydroxylase; TIM23: translocase of inner mitochondrial membrane 23; TOM20: translocase of outer mitochondrial membrane 20.

Techniques Used: shRNA, Injection, Expressing, Immunofluorescence, Fluorescence, Comparison, Membrane

Silencing PHB2 reduces antioxidative stress protein expression and aggravates motor defecits in PD mice. (A–D) Changes and quantitative analysis of Nrf2, HO-1, and NQO-1 protein levels in the midbrain of PHB2-shRNA and MPTP-treated PD mice. (E) Quantitative analysis of mice in the tail suspension and rotarod tests. Data are expressed as mean ± SEM ( n = 5). * P < 0.05, ** P < 0.01, vs. control group; $ P < 0.05, $$$ P < 0.001, vs . MPTP group (one-way analysis of variance with Tukey's multiple comparison test). HO-1: Heme oxygenase-1; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; PD: Parkinson's disease; PHB2: prohibitin 2.
Figure Legend Snippet: Silencing PHB2 reduces antioxidative stress protein expression and aggravates motor defecits in PD mice. (A–D) Changes and quantitative analysis of Nrf2, HO-1, and NQO-1 protein levels in the midbrain of PHB2-shRNA and MPTP-treated PD mice. (E) Quantitative analysis of mice in the tail suspension and rotarod tests. Data are expressed as mean ± SEM ( n = 5). * P < 0.05, ** P < 0.01, vs. control group; $ P < 0.05, $$$ P < 0.001, vs . MPTP group (one-way analysis of variance with Tukey's multiple comparison test). HO-1: Heme oxygenase-1; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; PD: Parkinson's disease; PHB2: prohibitin 2.

Techniques Used: Expressing, shRNA, Suspension, Comparison


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Proteintech antibodies against phb2
Antibodies Against Phb2, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Proteintech anti phb2
Anti Phb2, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Proteintech rabbit polyclonal anti phb2

Rabbit Polyclonal Anti Phb2, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit polyclonal anti phb2/product/Proteintech
Average 86 stars, based on 1 article reviews
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1) Product Images from "Identification of TMEM126A as OXA1L-interacting protein reveals cotranslational quality control in mitochondria"

Article Title: Identification of TMEM126A as OXA1L-interacting protein reveals cotranslational quality control in mitochondria

Journal: Molecular Cell

doi: 10.1016/j.molcel.2023.12.013


Figure Legend Snippet:

Techniques Used: Recombinant, Activity Assay, Software


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Proteintech anti phb2 mouse antibodies
Anti Phb2 Mouse Antibodies, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti phb2 mouse antibodies/product/Proteintech
Average 86 stars, based on 1 article reviews
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Proteintech anti phb2 mouse antibodies
<t>PHB2</t> upregulation is associated with poor patient outcomes in GC. A , B Representative images and quantitation of IHC analysis of PHB2 protein expression in 66 pairs of human gastric tumour tissues and corresponding adjacent normal gastric tissues. Scale bar, 50 μm. IRS: Immunoreactive score. Two-tailed Student’s t-test. C , D Western blotting ( C ) and qRT-PCR ( D ) analysis of PHB2 expression from 49 gastric tumour (T) patient samples and paired adjacent normal (N) gastric tissues. Data are representatives or mean ± SEM, two-tailed Student’s t-test. E PHB2 was upregulated in GC compared with normal gastric tissues as revealed by analysis of the STAD data in TCGA dataset. Data are mean ± SEM, two-tailed Student’s t-test. F The mRNA expression levels of PHB2 among different stages of GC tissues derived from the TCGA dataset. Data are mean ± SEM, one-way ANOVA followed by Tukey’s multiple comparison. G , H Western blotting ( G ) and qRT-PCR ( H ) analysis of PHB2 in a panel of GC cell and normal gastric epithelial cell line GES-1. Data are representatives or mean ± SEM; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison. I , J Kaplan–Meier analysis of the probability of overall survival of GC patients derived from the GEO datasets (GSE14210, GSE29272) using the optimal of PHB2 levels as the cut-off
Anti Phb2 Mouse Antibodies, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti phb2 mouse antibodies/product/Proteintech
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti phb2 mouse antibodies - by Bioz Stars, 2024-07
86/100 stars

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1) Product Images from "PHB2 promotes SHIP2 ubiquitination via the E3 ligase NEDD4 to regulate AKT signaling in gastric cancer"

Article Title: PHB2 promotes SHIP2 ubiquitination via the E3 ligase NEDD4 to regulate AKT signaling in gastric cancer

Journal: Journal of Experimental & Clinical Cancer Research : CR

doi: 10.1186/s13046-023-02937-1

PHB2 upregulation is associated with poor patient outcomes in GC. A , B Representative images and quantitation of IHC analysis of PHB2 protein expression in 66 pairs of human gastric tumour tissues and corresponding adjacent normal gastric tissues. Scale bar, 50 μm. IRS: Immunoreactive score. Two-tailed Student’s t-test. C , D Western blotting ( C ) and qRT-PCR ( D ) analysis of PHB2 expression from 49 gastric tumour (T) patient samples and paired adjacent normal (N) gastric tissues. Data are representatives or mean ± SEM, two-tailed Student’s t-test. E PHB2 was upregulated in GC compared with normal gastric tissues as revealed by analysis of the STAD data in TCGA dataset. Data are mean ± SEM, two-tailed Student’s t-test. F The mRNA expression levels of PHB2 among different stages of GC tissues derived from the TCGA dataset. Data are mean ± SEM, one-way ANOVA followed by Tukey’s multiple comparison. G , H Western blotting ( G ) and qRT-PCR ( H ) analysis of PHB2 in a panel of GC cell and normal gastric epithelial cell line GES-1. Data are representatives or mean ± SEM; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison. I , J Kaplan–Meier analysis of the probability of overall survival of GC patients derived from the GEO datasets (GSE14210, GSE29272) using the optimal of PHB2 levels as the cut-off
Figure Legend Snippet: PHB2 upregulation is associated with poor patient outcomes in GC. A , B Representative images and quantitation of IHC analysis of PHB2 protein expression in 66 pairs of human gastric tumour tissues and corresponding adjacent normal gastric tissues. Scale bar, 50 μm. IRS: Immunoreactive score. Two-tailed Student’s t-test. C , D Western blotting ( C ) and qRT-PCR ( D ) analysis of PHB2 expression from 49 gastric tumour (T) patient samples and paired adjacent normal (N) gastric tissues. Data are representatives or mean ± SEM, two-tailed Student’s t-test. E PHB2 was upregulated in GC compared with normal gastric tissues as revealed by analysis of the STAD data in TCGA dataset. Data are mean ± SEM, two-tailed Student’s t-test. F The mRNA expression levels of PHB2 among different stages of GC tissues derived from the TCGA dataset. Data are mean ± SEM, one-way ANOVA followed by Tukey’s multiple comparison. G , H Western blotting ( G ) and qRT-PCR ( H ) analysis of PHB2 in a panel of GC cell and normal gastric epithelial cell line GES-1. Data are representatives or mean ± SEM; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison. I , J Kaplan–Meier analysis of the probability of overall survival of GC patients derived from the GEO datasets (GSE14210, GSE29272) using the optimal of PHB2 levels as the cut-off

Techniques Used: Quantitation Assay, Expressing, Two Tailed Test, Western Blot, Quantitative RT-PCR, Derivative Assay, Comparison

PHB2 promotes GC cell proliferation and tumorigenicity. A-D shRNA silencing of PHB2 ( A ) attenuated GC cell proliferation as shown in MTT assays ( B ), clonogenic assays ( C ) and 5-bromo-2′-deoxyuridine (BrdU) incorporation ( D ). Data are representatives or mean ± SEM; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison. Scale bar, 1 cm. E , F PHB2 overexpression ( E ) promoted GC cell proliferation as shown in clonogenic assays ( F ). Data are representatives or mean ± SEM; n = 3 independent experiments, two-tailed Student’s t-test. Scale bar, 1 cm. G , H Overexpression of PHB2 ( G ) enhanced anchorage-independent growth of normal gastric epithelial cell line GES-1 ( H ). Data are representatives or mean ± SEM; n = 3 independent experiments, two-tailed Student’s t-test. Scale bar, 1 cm. I-K Representative photographs ( I ), tumour weight ( J ) and growth curves ( K ) of MKN-28.sh-scramble and MKN-28.sh-PHB2 xenografts in nu/nu mice. Data are representatives or mean ± SEM; n = 6 mice per group, two-tailed Student’s t-test ( J ), two-way ANOVA followed by Šídák’s multiple comparisons test ( K ). L Representative microscopic photographs (left panel), and quantitation of IHC staining for Ki67 (right panel) in FFPE sections of MKN-28.sh-scramble and MKN-28.sh-PHB2 xenografts. Data are representatives or mean ± SEM; n = 6 mice per group, two-tailed Student’s t-test. Scale bar, 20 μm. IRS: Immunoreactive score
Figure Legend Snippet: PHB2 promotes GC cell proliferation and tumorigenicity. A-D shRNA silencing of PHB2 ( A ) attenuated GC cell proliferation as shown in MTT assays ( B ), clonogenic assays ( C ) and 5-bromo-2′-deoxyuridine (BrdU) incorporation ( D ). Data are representatives or mean ± SEM; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison. Scale bar, 1 cm. E , F PHB2 overexpression ( E ) promoted GC cell proliferation as shown in clonogenic assays ( F ). Data are representatives or mean ± SEM; n = 3 independent experiments, two-tailed Student’s t-test. Scale bar, 1 cm. G , H Overexpression of PHB2 ( G ) enhanced anchorage-independent growth of normal gastric epithelial cell line GES-1 ( H ). Data are representatives or mean ± SEM; n = 3 independent experiments, two-tailed Student’s t-test. Scale bar, 1 cm. I-K Representative photographs ( I ), tumour weight ( J ) and growth curves ( K ) of MKN-28.sh-scramble and MKN-28.sh-PHB2 xenografts in nu/nu mice. Data are representatives or mean ± SEM; n = 6 mice per group, two-tailed Student’s t-test ( J ), two-way ANOVA followed by Šídák’s multiple comparisons test ( K ). L Representative microscopic photographs (left panel), and quantitation of IHC staining for Ki67 (right panel) in FFPE sections of MKN-28.sh-scramble and MKN-28.sh-PHB2 xenografts. Data are representatives or mean ± SEM; n = 6 mice per group, two-tailed Student’s t-test. Scale bar, 20 μm. IRS: Immunoreactive score

Techniques Used: shRNA, BrdU Incorporation Assay, Comparison, Over Expression, Two Tailed Test, Quantitation Assay, Immunohistochemistry

PHB2 binds to and co-localizes with SHIP2 in the cytoplasm of GC cells. A Endogenous PHB2 and SHIP2 were co-precipitated with each other in HGC-27 and MKN-28 cells. B Exogenous Flag-PHB2, and tGFP-SHIP2 were co-precipitated with each other in AGS cells. C Representative immunofluorescence photographs of PHB2 and SHIP2 co-localization in HGC-27 cells. Scale bar, 10 μm. D Western blotting showing subcellular localization of PHB2 and SHIP2. Cyt: cytoplasm; Nuc: nucleus. β-actin: Cyt marker; Histone H3: Nuc marker. E tGFP-SHIP2 was co-pulled down by recombinant GST-PHB1 or GST-PHB2, which was however diminished by PHB2 knockdown. Data are representatives of three independent experiments
Figure Legend Snippet: PHB2 binds to and co-localizes with SHIP2 in the cytoplasm of GC cells. A Endogenous PHB2 and SHIP2 were co-precipitated with each other in HGC-27 and MKN-28 cells. B Exogenous Flag-PHB2, and tGFP-SHIP2 were co-precipitated with each other in AGS cells. C Representative immunofluorescence photographs of PHB2 and SHIP2 co-localization in HGC-27 cells. Scale bar, 10 μm. D Western blotting showing subcellular localization of PHB2 and SHIP2. Cyt: cytoplasm; Nuc: nucleus. β-actin: Cyt marker; Histone H3: Nuc marker. E tGFP-SHIP2 was co-pulled down by recombinant GST-PHB1 or GST-PHB2, which was however diminished by PHB2 knockdown. Data are representatives of three independent experiments

Techniques Used: Immunofluorescence, Western Blot, Marker, Recombinant

The SAM domain of SHIP2 and all regions of PHB2 except for the N-terminal are required for the interaction between PHB2 and SHIP2. A Schematic illustration of full-length SHIP2 (SHIP2-FL) and its corresponding truncates used in mapping experiments. B Deletion-mapping experiments showing that the SAM domain of SHIP2 is indispensable for its interaction with PHB2 in HEK293T cells. C Schematic illustration of full-length PHB2 (PHB2-FL) and the PHB2 mutants. D Deletion-mapping experiments showing that the N-terminal region of PHB2 is dispensable for its interaction with SHIP2 in HEK293T cells. Data are representatives of three independent experiments
Figure Legend Snippet: The SAM domain of SHIP2 and all regions of PHB2 except for the N-terminal are required for the interaction between PHB2 and SHIP2. A Schematic illustration of full-length SHIP2 (SHIP2-FL) and its corresponding truncates used in mapping experiments. B Deletion-mapping experiments showing that the SAM domain of SHIP2 is indispensable for its interaction with PHB2 in HEK293T cells. C Schematic illustration of full-length PHB2 (PHB2-FL) and the PHB2 mutants. D Deletion-mapping experiments showing that the N-terminal region of PHB2 is dispensable for its interaction with SHIP2 in HEK293T cells. Data are representatives of three independent experiments

Techniques Used:

PHB2 promotes SHIP2 protein degradation through ubiquitination. A , B Western blotting ( A ) and qRT-PCR analysis ( B ) of SHIP2 expression in HGC-27.sh-PHB2 and MKN-28.sh-PHB2 cells. Data are representatives or mean ± SEM; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison. C Silencing of PHB2 prolonged the half-life time of SHIP2 protein in cycloheximide (CHX)-chase assays. Data are representatives or mean ± SEM; n = 3 independent experiments, two-way ANOVA followed by Šídák’s multiple comparisons test. CHX: 10 μg/ml. D Silencing of PHB2 reduced overexpressed exogenous SHIP2 ubiquitination. MG132:10 μM. Data shown represent three independent experiments. E Overexpression of PHB2 enhanced the ubiquitination of exogenous full-length SHIP2 but not tGFP-SHIP2-del SAM mutant SHIP2. MG132: 10 μM. Data shown represent three independent experiments
Figure Legend Snippet: PHB2 promotes SHIP2 protein degradation through ubiquitination. A , B Western blotting ( A ) and qRT-PCR analysis ( B ) of SHIP2 expression in HGC-27.sh-PHB2 and MKN-28.sh-PHB2 cells. Data are representatives or mean ± SEM; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison. C Silencing of PHB2 prolonged the half-life time of SHIP2 protein in cycloheximide (CHX)-chase assays. Data are representatives or mean ± SEM; n = 3 independent experiments, two-way ANOVA followed by Šídák’s multiple comparisons test. CHX: 10 μg/ml. D Silencing of PHB2 reduced overexpressed exogenous SHIP2 ubiquitination. MG132:10 μM. Data shown represent three independent experiments. E Overexpression of PHB2 enhanced the ubiquitination of exogenous full-length SHIP2 but not tGFP-SHIP2-del SAM mutant SHIP2. MG132: 10 μM. Data shown represent three independent experiments

Techniques Used: Western Blot, Quantitative RT-PCR, Expressing, Comparison, Over Expression, Mutagenesis

PHB2 induces the ubiquitination degradation of SHIP2 by enhancing the interaction between NEDD4 and SHIP2. A Co-IP assays showing that NEDD4 was specifically co-precipitated with SHIP2 and PHB2 in HGC-27 cells. E3 ligases SIAH2, WWP2, MUL1 and Cul4A were used as negative controls. Data shown represent three independent experiments. B SiRNA knockdown of NEDD4 decreased the ubiquitination of SHIP2. MG132: 10 μM. C Deletion-mapping experiments showing that NEDD4 was precipitated with the 5-Ptase domain of SHIP2 but not other SHIP2 domains in HEK293T cells. D Overexpression of NEDD4 increased the ubiquitination full-length SHIP2 but not tGFP-SHIP2-del 5Ptase in HEK293T and HGC-27 cells. MG132:10 μM. E Silencing of PHB2 reduced the interaction between SHIP2 and NEDD4 in HEK293T cells. F Silencing of NEDD4 abolished the ubiquitination of SHIP2 caused by PHB2 overexpression. G, Overexpression of PHB2 mainly increased K48-linked polyubiquitination of SHIP2. MG132:10 μM. Data are representatives of three independent experiments
Figure Legend Snippet: PHB2 induces the ubiquitination degradation of SHIP2 by enhancing the interaction between NEDD4 and SHIP2. A Co-IP assays showing that NEDD4 was specifically co-precipitated with SHIP2 and PHB2 in HGC-27 cells. E3 ligases SIAH2, WWP2, MUL1 and Cul4A were used as negative controls. Data shown represent three independent experiments. B SiRNA knockdown of NEDD4 decreased the ubiquitination of SHIP2. MG132: 10 μM. C Deletion-mapping experiments showing that NEDD4 was precipitated with the 5-Ptase domain of SHIP2 but not other SHIP2 domains in HEK293T cells. D Overexpression of NEDD4 increased the ubiquitination full-length SHIP2 but not tGFP-SHIP2-del 5Ptase in HEK293T and HGC-27 cells. MG132:10 μM. E Silencing of PHB2 reduced the interaction between SHIP2 and NEDD4 in HEK293T cells. F Silencing of NEDD4 abolished the ubiquitination of SHIP2 caused by PHB2 overexpression. G, Overexpression of PHB2 mainly increased K48-linked polyubiquitination of SHIP2. MG132:10 μM. Data are representatives of three independent experiments

Techniques Used: Co-Immunoprecipitation Assay, Over Expression

PHB2-mediated SHIP2 degradation leads to Akt activation and GC proliferation. A Knockdown of PHB2 increased SHIP2 protein expression and diminished Akt activation, which was reversed by co-knockdown of SHIP2. Data shown represent three independent experiments. B , C Knockdown of PHB2 inhibited GC cell proliferation, which was abolished by co-knockdown of SHIP2 as shown in clonogenic assays ( B ) and BrdU incorporation ( C ). Data are representatives or mean ± SEM; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison. Scale bar, 1 cm. D-F, Overexpression of myr-Akt reversed the inhibition of Akt signaling ( D ) and GC cell proliferation caused by silencing of PHB2 as shown in clonogenic assays ( E ) and BrdU incorporation ( F ). Data are representatives or mean ± SEM; n = 3 independent experiments, two-tailed Student’s t-test. Scale bar, 1 cm. G-I Representative Photographs ( G ), tumour weight ( H ) and growth curves ( I ) of MKN-28.sh-scramble and MKN-28.sh-PHB2 xenografts in nu/nu mice with or without co-transduction of myr-Akt. Data are representatives or mean ± SEM; n = 3 mice per group, one-way ANOVA followed by Tukey’s multiple comparison ( H ), two-way ANOVA followed by Tukey’s multiple comparison ( I ). J Western blotting showing the expression of PHB2, SHIP2 and Akt in GC cells isolated from MKN-28.sh-scramble and MKN-28.sh-PHB2 xenografts harvested from mice treated as described in G . Data are representatives of three independent experiments
Figure Legend Snippet: PHB2-mediated SHIP2 degradation leads to Akt activation and GC proliferation. A Knockdown of PHB2 increased SHIP2 protein expression and diminished Akt activation, which was reversed by co-knockdown of SHIP2. Data shown represent three independent experiments. B , C Knockdown of PHB2 inhibited GC cell proliferation, which was abolished by co-knockdown of SHIP2 as shown in clonogenic assays ( B ) and BrdU incorporation ( C ). Data are representatives or mean ± SEM; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison. Scale bar, 1 cm. D-F, Overexpression of myr-Akt reversed the inhibition of Akt signaling ( D ) and GC cell proliferation caused by silencing of PHB2 as shown in clonogenic assays ( E ) and BrdU incorporation ( F ). Data are representatives or mean ± SEM; n = 3 independent experiments, two-tailed Student’s t-test. Scale bar, 1 cm. G-I Representative Photographs ( G ), tumour weight ( H ) and growth curves ( I ) of MKN-28.sh-scramble and MKN-28.sh-PHB2 xenografts in nu/nu mice with or without co-transduction of myr-Akt. Data are representatives or mean ± SEM; n = 3 mice per group, one-way ANOVA followed by Tukey’s multiple comparison ( H ), two-way ANOVA followed by Tukey’s multiple comparison ( I ). J Western blotting showing the expression of PHB2, SHIP2 and Akt in GC cells isolated from MKN-28.sh-scramble and MKN-28.sh-PHB2 xenografts harvested from mice treated as described in G . Data are representatives of three independent experiments

Techniques Used: Activation Assay, Expressing, BrdU Incorporation Assay, Comparison, Over Expression, Inhibition, Two Tailed Test, Transduction, Western Blot, Isolation

PHB2 expression is negatively correlated with SHIP2 expression, and positively correlated with p-Akt and Ki67 expression in GC. A representative photograph showed the expression of PHB2, SHIP2, p-Akt, and Ki67 in GC tissues and adjacent normal tissues examined by IHC staining ( n = 15). Scale bar: 200 μm. B-D the expression correlations between PHB2 and SHIP2 ( B ), PHB2 and p-Akt ( C ), and PHB2 and Ki67 ( D ) depicted in A were analysed using using Pearson’s correlation coefficient test. n = 15 clinical samples. E , F the expression correlations between PHB2 and SHIP2 ( E ), and PHB2 and Ki67 ( F ) in GC tissues from the Human Protein Atlas dataset ( https://www.proteinatlas.org ) were analysed using Pearson’s correlation coefficient test. n = 9 clinical samples. G Western blotting analysis of PHB2, SHIP2, p-Akt, and GAPDH protein expression in gastric tumour (T) patient samples and paired adjacent normal (N) gastric tissues. Data are representatives
Figure Legend Snippet: PHB2 expression is negatively correlated with SHIP2 expression, and positively correlated with p-Akt and Ki67 expression in GC. A representative photograph showed the expression of PHB2, SHIP2, p-Akt, and Ki67 in GC tissues and adjacent normal tissues examined by IHC staining ( n = 15). Scale bar: 200 μm. B-D the expression correlations between PHB2 and SHIP2 ( B ), PHB2 and p-Akt ( C ), and PHB2 and Ki67 ( D ) depicted in A were analysed using using Pearson’s correlation coefficient test. n = 15 clinical samples. E , F the expression correlations between PHB2 and SHIP2 ( E ), and PHB2 and Ki67 ( F ) in GC tissues from the Human Protein Atlas dataset ( https://www.proteinatlas.org ) were analysed using Pearson’s correlation coefficient test. n = 9 clinical samples. G Western blotting analysis of PHB2, SHIP2, p-Akt, and GAPDH protein expression in gastric tumour (T) patient samples and paired adjacent normal (N) gastric tissues. Data are representatives

Techniques Used: Expressing, Immunohistochemistry, Western Blot

Schematic illustrating the role of PHB2 in regulating GC tumorigenesis. SHIP2 acts as a negative regulator of phosphoinositide 3-kinase (PI3K) and insulin signaling. In GC cells, high expression of PHB2 enhanced the interaction between NEDD4 and SHIP2, which triggers the ubiquitination degradation of SHIP2, consequently leading to the activation of Akt and promoting GC proliferation and tumorigenesis
Figure Legend Snippet: Schematic illustrating the role of PHB2 in regulating GC tumorigenesis. SHIP2 acts as a negative regulator of phosphoinositide 3-kinase (PI3K) and insulin signaling. In GC cells, high expression of PHB2 enhanced the interaction between NEDD4 and SHIP2, which triggers the ubiquitination degradation of SHIP2, consequently leading to the activation of Akt and promoting GC proliferation and tumorigenesis

Techniques Used: Expressing, Activation Assay


Structured Review

Proteintech anti phb2
Anti Phb2, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 86 stars, based on 1 article reviews
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anti phb2 - by Bioz Stars, 2024-07
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Structured Review

Proteintech rabbit anti phb2
APOL1- and APOL3-specific interactions (A) Interaction of APOL1, APOL1 mutants, or APOL3 with <t>PHB2,</t> as measured in the E. coli pDUET co-expression system. Nickel binding of S-tagged proteins (yellow frame) reflects their relative association with His-tagged partners (green frame). These data are representative of at least 3 independent experiments. A1, WT APOL1; Δ, APOL1Δ; A3, WT APOL3; H4Q, APOL1 mutHel4Q; H5Q, APOL1 mutHel5Q; H5P, APOL1 mutHel5P; H5mS, APOL1 mutHel5S; H5sw, APOL1 Hel5 swap; BH3E, APOL1 BH3 mutant E. (B) Representative SPR sensorgrams of immobilized L1Hel5 (left) or L3Hel5 (right) peptide interaction with recombinant PHB2 with or without calcium (n = 3). (C) Representative SPR sensorgrams of immobilized L1Hel5 (left) or L3Hel5 (right) peptide interaction with APOL3 Hel5, NTD, or CTD peptide (n = 3). (D) SPR measurements of bound sVAMP8 (left) or TbVAMP7Bp (right) interaction with various APOL3 versions as indicated (n = 3). (E) SPR measurements of bound sVAMP8 (left) or TbVAMP7Bp (right) interaction with different APOL3 peptides as indicated (n = 3). (F) Immunodetection of recombinant sVAMP8 binding to various lipids spotted on membrane strips. LPA, lysophosphatidic acid; LPC, lysophosphocholine; PI, phosphatidylinositol; PE, phosphatidylethanolamine; PC, phosphatidylcholine; S1P, sphingosine-1-phosphate; PA, phosphatidic acid; PS, phosphatidylserine; TG, triglyceride; DAG, diacylglycerol; PG, phosphatidylglycerol; CL, cardiolipin; Chol, cholesterol; SM, sphingomyelin. Blank, no lipid.
Rabbit Anti Phb2, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit anti phb2/product/Proteintech
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
rabbit anti phb2 - by Bioz Stars, 2024-07
86/100 stars

Images

1) Product Images from "Apolipoproteins L1 and L3 control mitochondrial membrane dynamics"

Article Title: Apolipoproteins L1 and L3 control mitochondrial membrane dynamics

Journal: Cell Reports

doi: 10.1016/j.celrep.2023.113528

APOL1- and APOL3-specific interactions (A) Interaction of APOL1, APOL1 mutants, or APOL3 with PHB2, as measured in the E. coli pDUET co-expression system. Nickel binding of S-tagged proteins (yellow frame) reflects their relative association with His-tagged partners (green frame). These data are representative of at least 3 independent experiments. A1, WT APOL1; Δ, APOL1Δ; A3, WT APOL3; H4Q, APOL1 mutHel4Q; H5Q, APOL1 mutHel5Q; H5P, APOL1 mutHel5P; H5mS, APOL1 mutHel5S; H5sw, APOL1 Hel5 swap; BH3E, APOL1 BH3 mutant E. (B) Representative SPR sensorgrams of immobilized L1Hel5 (left) or L3Hel5 (right) peptide interaction with recombinant PHB2 with or without calcium (n = 3). (C) Representative SPR sensorgrams of immobilized L1Hel5 (left) or L3Hel5 (right) peptide interaction with APOL3 Hel5, NTD, or CTD peptide (n = 3). (D) SPR measurements of bound sVAMP8 (left) or TbVAMP7Bp (right) interaction with various APOL3 versions as indicated (n = 3). (E) SPR measurements of bound sVAMP8 (left) or TbVAMP7Bp (right) interaction with different APOL3 peptides as indicated (n = 3). (F) Immunodetection of recombinant sVAMP8 binding to various lipids spotted on membrane strips. LPA, lysophosphatidic acid; LPC, lysophosphocholine; PI, phosphatidylinositol; PE, phosphatidylethanolamine; PC, phosphatidylcholine; S1P, sphingosine-1-phosphate; PA, phosphatidic acid; PS, phosphatidylserine; TG, triglyceride; DAG, diacylglycerol; PG, phosphatidylglycerol; CL, cardiolipin; Chol, cholesterol; SM, sphingomyelin. Blank, no lipid.
Figure Legend Snippet: APOL1- and APOL3-specific interactions (A) Interaction of APOL1, APOL1 mutants, or APOL3 with PHB2, as measured in the E. coli pDUET co-expression system. Nickel binding of S-tagged proteins (yellow frame) reflects their relative association with His-tagged partners (green frame). These data are representative of at least 3 independent experiments. A1, WT APOL1; Δ, APOL1Δ; A3, WT APOL3; H4Q, APOL1 mutHel4Q; H5Q, APOL1 mutHel5Q; H5P, APOL1 mutHel5P; H5mS, APOL1 mutHel5S; H5sw, APOL1 Hel5 swap; BH3E, APOL1 BH3 mutant E. (B) Representative SPR sensorgrams of immobilized L1Hel5 (left) or L3Hel5 (right) peptide interaction with recombinant PHB2 with or without calcium (n = 3). (C) Representative SPR sensorgrams of immobilized L1Hel5 (left) or L3Hel5 (right) peptide interaction with APOL3 Hel5, NTD, or CTD peptide (n = 3). (D) SPR measurements of bound sVAMP8 (left) or TbVAMP7Bp (right) interaction with various APOL3 versions as indicated (n = 3). (E) SPR measurements of bound sVAMP8 (left) or TbVAMP7Bp (right) interaction with different APOL3 peptides as indicated (n = 3). (F) Immunodetection of recombinant sVAMP8 binding to various lipids spotted on membrane strips. LPA, lysophosphatidic acid; LPC, lysophosphocholine; PI, phosphatidylinositol; PE, phosphatidylethanolamine; PC, phosphatidylcholine; S1P, sphingosine-1-phosphate; PA, phosphatidic acid; PS, phosphatidylserine; TG, triglyceride; DAG, diacylglycerol; PG, phosphatidylglycerol; CL, cardiolipin; Chol, cholesterol; SM, sphingomyelin. Blank, no lipid.

Techniques Used: Expressing, Binding Assay, Mutagenesis, Recombinant, Immunodetection, Membrane


Figure Legend Snippet:

Techniques Used: Recombinant, Kinase Assay, Stripping Membranes, Software

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    Proteintech phb2
    Phb2, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech antibodies against phb2
    Effect of MPP + on <t>PHB2</t> protein expression and mitochondrial function in SH-SY5Y cells. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp, and then induced with 1 mM MPP + for 24 hours. (A, B) PHB2 protein concentration in SH-SY5Y cells after 1 mM MPP + treatment for 6, 12, 24, and 48 hours. (A) Western blots showing PHB2 protein expression in MPP + -treated cells at different times. (B) Quantitative analysis of PHB2 protein expression. (C–G) Protein expression changes of Nrf2, HO-1, NQO-1, and PHB2. (C) Western blots showing Nrf2, HO-1, NQO-1, and PHB2 protein expression in MPP + -treated and PHB2-shRNA- or PHB2-Over Exp-treated cells. (D–G) Quantitative analysis of Nrf2, HO-1, NQO-1, and PHB2 protein expression. (H, I) The effect of PHB2 expression on mitochondrial membrane potential induced by MPP + . (H) JC-1 staining of SH-SY5Y cells treated with PHB2-shRNA and PHB2-Over Exp. Original magnification 10×, scale bar: 50 μm. (I) Quantitative analysis of JC-1 staining: the green/red ratio reflects changes in the mitochondrial membrane potential. (J, K) The effect of changes in PHB2 expression on ROS production under MPP + induction. (J) Intracellular ROS fluorescence. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp and labeled with DCFH-DA (green fluorescence). Original magnification 4×, scale bar: 100 μm. (K) Quantitative analysis of ROS (DCFH-DA) in SH-SY5Y cells. (L) Electron microscopy images of morphological changes in mitochondria in PHB2-shRNA-treated and MPP + -induced cells. Red boxes show magnified mitochondria. Scale bars: 1 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . control group; # P < 0.05,## P < 0.01, vs . MPP + group (one-way analysis of variance with Tukey's multiple comparison test). DCFH-DA: 2′,7′-Dichlorodihydrofluorescein diacetate; HO-1: heme oxygenase-1; JC-1: 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide; MPP + : 1-methyl-4-phenylpyridinium; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; Over Exp: overexpression; PHB2: prohibitin 2; ROS: reactive oxygen species; SH-SY5Y: human neuroblastoma cell line.
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    Proteintech anti phb2
    Effect of MPP + on <t>PHB2</t> protein expression and mitochondrial function in SH-SY5Y cells. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp, and then induced with 1 mM MPP + for 24 hours. (A, B) PHB2 protein concentration in SH-SY5Y cells after 1 mM MPP + treatment for 6, 12, 24, and 48 hours. (A) Western blots showing PHB2 protein expression in MPP + -treated cells at different times. (B) Quantitative analysis of PHB2 protein expression. (C–G) Protein expression changes of Nrf2, HO-1, NQO-1, and PHB2. (C) Western blots showing Nrf2, HO-1, NQO-1, and PHB2 protein expression in MPP + -treated and PHB2-shRNA- or PHB2-Over Exp-treated cells. (D–G) Quantitative analysis of Nrf2, HO-1, NQO-1, and PHB2 protein expression. (H, I) The effect of PHB2 expression on mitochondrial membrane potential induced by MPP + . (H) JC-1 staining of SH-SY5Y cells treated with PHB2-shRNA and PHB2-Over Exp. Original magnification 10×, scale bar: 50 μm. (I) Quantitative analysis of JC-1 staining: the green/red ratio reflects changes in the mitochondrial membrane potential. (J, K) The effect of changes in PHB2 expression on ROS production under MPP + induction. (J) Intracellular ROS fluorescence. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp and labeled with DCFH-DA (green fluorescence). Original magnification 4×, scale bar: 100 μm. (K) Quantitative analysis of ROS (DCFH-DA) in SH-SY5Y cells. (L) Electron microscopy images of morphological changes in mitochondria in PHB2-shRNA-treated and MPP + -induced cells. Red boxes show magnified mitochondria. Scale bars: 1 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . control group; # P < 0.05,## P < 0.01, vs . MPP + group (one-way analysis of variance with Tukey's multiple comparison test). DCFH-DA: 2′,7′-Dichlorodihydrofluorescein diacetate; HO-1: heme oxygenase-1; JC-1: 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide; MPP + : 1-methyl-4-phenylpyridinium; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; Over Exp: overexpression; PHB2: prohibitin 2; ROS: reactive oxygen species; SH-SY5Y: human neuroblastoma cell line.
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    86
    Proteintech rabbit anti phb2
    APOL1- and APOL3-specific interactions (A) Interaction of APOL1, APOL1 mutants, or APOL3 with <t>PHB2,</t> as measured in the E. coli pDUET co-expression system. Nickel binding of S-tagged proteins (yellow frame) reflects their relative association with His-tagged partners (green frame). These data are representative of at least 3 independent experiments. A1, WT APOL1; Δ, APOL1Δ; A3, WT APOL3; H4Q, APOL1 mutHel4Q; H5Q, APOL1 mutHel5Q; H5P, APOL1 mutHel5P; H5mS, APOL1 mutHel5S; H5sw, APOL1 Hel5 swap; BH3E, APOL1 BH3 mutant E. (B) Representative SPR sensorgrams of immobilized L1Hel5 (left) or L3Hel5 (right) peptide interaction with recombinant PHB2 with or without calcium (n = 3). (C) Representative SPR sensorgrams of immobilized L1Hel5 (left) or L3Hel5 (right) peptide interaction with APOL3 Hel5, NTD, or CTD peptide (n = 3). (D) SPR measurements of bound sVAMP8 (left) or TbVAMP7Bp (right) interaction with various APOL3 versions as indicated (n = 3). (E) SPR measurements of bound sVAMP8 (left) or TbVAMP7Bp (right) interaction with different APOL3 peptides as indicated (n = 3). (F) Immunodetection of recombinant sVAMP8 binding to various lipids spotted on membrane strips. LPA, lysophosphatidic acid; LPC, lysophosphocholine; PI, phosphatidylinositol; PE, phosphatidylethanolamine; PC, phosphatidylcholine; S1P, sphingosine-1-phosphate; PA, phosphatidic acid; PS, phosphatidylserine; TG, triglyceride; DAG, diacylglycerol; PG, phosphatidylglycerol; CL, cardiolipin; Chol, cholesterol; SM, sphingomyelin. Blank, no lipid.
    Rabbit Anti Phb2, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effect of MPP + on PHB2 protein expression and mitochondrial function in SH-SY5Y cells. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp, and then induced with 1 mM MPP + for 24 hours. (A, B) PHB2 protein concentration in SH-SY5Y cells after 1 mM MPP + treatment for 6, 12, 24, and 48 hours. (A) Western blots showing PHB2 protein expression in MPP + -treated cells at different times. (B) Quantitative analysis of PHB2 protein expression. (C–G) Protein expression changes of Nrf2, HO-1, NQO-1, and PHB2. (C) Western blots showing Nrf2, HO-1, NQO-1, and PHB2 protein expression in MPP + -treated and PHB2-shRNA- or PHB2-Over Exp-treated cells. (D–G) Quantitative analysis of Nrf2, HO-1, NQO-1, and PHB2 protein expression. (H, I) The effect of PHB2 expression on mitochondrial membrane potential induced by MPP + . (H) JC-1 staining of SH-SY5Y cells treated with PHB2-shRNA and PHB2-Over Exp. Original magnification 10×, scale bar: 50 μm. (I) Quantitative analysis of JC-1 staining: the green/red ratio reflects changes in the mitochondrial membrane potential. (J, K) The effect of changes in PHB2 expression on ROS production under MPP + induction. (J) Intracellular ROS fluorescence. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp and labeled with DCFH-DA (green fluorescence). Original magnification 4×, scale bar: 100 μm. (K) Quantitative analysis of ROS (DCFH-DA) in SH-SY5Y cells. (L) Electron microscopy images of morphological changes in mitochondria in PHB2-shRNA-treated and MPP + -induced cells. Red boxes show magnified mitochondria. Scale bars: 1 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . control group; # P < 0.05,## P < 0.01, vs . MPP + group (one-way analysis of variance with Tukey's multiple comparison test). DCFH-DA: 2′,7′-Dichlorodihydrofluorescein diacetate; HO-1: heme oxygenase-1; JC-1: 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide; MPP + : 1-methyl-4-phenylpyridinium; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; Over Exp: overexpression; PHB2: prohibitin 2; ROS: reactive oxygen species; SH-SY5Y: human neuroblastoma cell line.

    Journal: Neural Regeneration Research

    Article Title: A novel mechanism of PHB2-mediated mitophagy participating in the development of Parkinson's disease

    doi: 10.4103/1673-5374.389356

    Figure Lengend Snippet: Effect of MPP + on PHB2 protein expression and mitochondrial function in SH-SY5Y cells. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp, and then induced with 1 mM MPP + for 24 hours. (A, B) PHB2 protein concentration in SH-SY5Y cells after 1 mM MPP + treatment for 6, 12, 24, and 48 hours. (A) Western blots showing PHB2 protein expression in MPP + -treated cells at different times. (B) Quantitative analysis of PHB2 protein expression. (C–G) Protein expression changes of Nrf2, HO-1, NQO-1, and PHB2. (C) Western blots showing Nrf2, HO-1, NQO-1, and PHB2 protein expression in MPP + -treated and PHB2-shRNA- or PHB2-Over Exp-treated cells. (D–G) Quantitative analysis of Nrf2, HO-1, NQO-1, and PHB2 protein expression. (H, I) The effect of PHB2 expression on mitochondrial membrane potential induced by MPP + . (H) JC-1 staining of SH-SY5Y cells treated with PHB2-shRNA and PHB2-Over Exp. Original magnification 10×, scale bar: 50 μm. (I) Quantitative analysis of JC-1 staining: the green/red ratio reflects changes in the mitochondrial membrane potential. (J, K) The effect of changes in PHB2 expression on ROS production under MPP + induction. (J) Intracellular ROS fluorescence. SH-SY5Y cells were treated with PHB2-shRNA and PHB2-Over Exp and labeled with DCFH-DA (green fluorescence). Original magnification 4×, scale bar: 100 μm. (K) Quantitative analysis of ROS (DCFH-DA) in SH-SY5Y cells. (L) Electron microscopy images of morphological changes in mitochondria in PHB2-shRNA-treated and MPP + -induced cells. Red boxes show magnified mitochondria. Scale bars: 1 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . control group; # P < 0.05,## P < 0.01, vs . MPP + group (one-way analysis of variance with Tukey's multiple comparison test). DCFH-DA: 2′,7′-Dichlorodihydrofluorescein diacetate; HO-1: heme oxygenase-1; JC-1: 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide; MPP + : 1-methyl-4-phenylpyridinium; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; Over Exp: overexpression; PHB2: prohibitin 2; ROS: reactive oxygen species; SH-SY5Y: human neuroblastoma cell line.

    Article Snippet: Membranes were incubated overnight at 4°C in solutions with primary antibodies against PHB2 (rabbit, 1:2000, Proteintech, Chicago, IL, USA, Cat# 12295-1-AP, RRID: AB_2164779), Parkin (mouse, 1:2000, Abcam, Cambridge, UK, Cat# ab77924, RRID: AB_1566559), LC3B (rabbit, 1:2000, Abcam, Cat# ab192890, RRID: AB_2827794), p-PERK Thr980 (rabbit, 1:2000, Cell Signaling Technology, Danvers, MA, USA, Cat# 3179, RRID: AB_2095853), nuclear factor erythroid 2-related factor (Nrf2; rabbit, 1:2000, Cell Signaling Technology, Cat# 12721, RRID: AB_2715528), heme oxygenase 1 (HO-1; rabbit, 1:2000, Cell Signaling Technology, Cat# 43966, RRID: AB_2799254), quinone oxidoreductase 1 (NQO-1; mouse, 1:2000, Cell Signaling Technology, Cat #3187, RRID: AB_2154354), and β-actin (mouse, 1:5000, CWBio, Beijing, China, Cat# CW0096, RRID: AB_2665433).

    Techniques: Expressing, shRNA, Protein Concentration, Western Blot, Membrane, Staining, Fluorescence, Labeling, Electron Microscopy, Comparison, Over Expression

    Effect of PHB2 on mitophagy in a PD cell model. (A–F) PHB2 and LC3II/LC3I protein expression in SH-SY5Y cells after treatment with PHB2-shRNA or PHB2-Over Exp and MPP + (1 mM, 24 hours). (A) Protein expression levels of PHB2 and LC3 in SH-SY5Y cells treated with PHB2-shRNA under MPP + induction. (B, C) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (D) Protein expression levels of PHB2 and LC3 in SH-SY5Y cells treated with PHB2-Over Exp under MPP + induction. (E, F) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (G) Cellular localization of LC3 and TOM20 or TIM23 in SH-SY5Y cells treated with PHB2-shRNA under MPP + induction. Immunofluorescence changes of LC3 and mitochondrial proteins (TOM20 and TIM23). Red fluorescence represents TOM20 and TIM23, green fluorescence represents LC3, and blue fluorescence represents DAPI. Original magnification 20×, Scale bars: 20 μm. Data are expressed as mean ± SEM ( n = 3). ** P < 0.01, *** P < 0.001, vs . control group (one-way analysis of variance with Tukey's multiple comparison test). DAPI: 4′,6-Diamidino-2-phenylindole; LC3: microtubule-associated protein 1 light chain 3; MPP + : 1-methyl-4-phenylpyridinium; Over Exp: overexpression; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line; TIM23: translocase of inner mitochondrial membrane 23; TOM20: translocase of outer mitochondrial membrane 20.

    Journal: Neural Regeneration Research

    Article Title: A novel mechanism of PHB2-mediated mitophagy participating in the development of Parkinson's disease

    doi: 10.4103/1673-5374.389356

    Figure Lengend Snippet: Effect of PHB2 on mitophagy in a PD cell model. (A–F) PHB2 and LC3II/LC3I protein expression in SH-SY5Y cells after treatment with PHB2-shRNA or PHB2-Over Exp and MPP + (1 mM, 24 hours). (A) Protein expression levels of PHB2 and LC3 in SH-SY5Y cells treated with PHB2-shRNA under MPP + induction. (B, C) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (D) Protein expression levels of PHB2 and LC3 in SH-SY5Y cells treated with PHB2-Over Exp under MPP + induction. (E, F) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (G) Cellular localization of LC3 and TOM20 or TIM23 in SH-SY5Y cells treated with PHB2-shRNA under MPP + induction. Immunofluorescence changes of LC3 and mitochondrial proteins (TOM20 and TIM23). Red fluorescence represents TOM20 and TIM23, green fluorescence represents LC3, and blue fluorescence represents DAPI. Original magnification 20×, Scale bars: 20 μm. Data are expressed as mean ± SEM ( n = 3). ** P < 0.01, *** P < 0.001, vs . control group (one-way analysis of variance with Tukey's multiple comparison test). DAPI: 4′,6-Diamidino-2-phenylindole; LC3: microtubule-associated protein 1 light chain 3; MPP + : 1-methyl-4-phenylpyridinium; Over Exp: overexpression; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line; TIM23: translocase of inner mitochondrial membrane 23; TOM20: translocase of outer mitochondrial membrane 20.

    Article Snippet: Membranes were incubated overnight at 4°C in solutions with primary antibodies against PHB2 (rabbit, 1:2000, Proteintech, Chicago, IL, USA, Cat# 12295-1-AP, RRID: AB_2164779), Parkin (mouse, 1:2000, Abcam, Cambridge, UK, Cat# ab77924, RRID: AB_1566559), LC3B (rabbit, 1:2000, Abcam, Cat# ab192890, RRID: AB_2827794), p-PERK Thr980 (rabbit, 1:2000, Cell Signaling Technology, Danvers, MA, USA, Cat# 3179, RRID: AB_2095853), nuclear factor erythroid 2-related factor (Nrf2; rabbit, 1:2000, Cell Signaling Technology, Cat# 12721, RRID: AB_2715528), heme oxygenase 1 (HO-1; rabbit, 1:2000, Cell Signaling Technology, Cat# 43966, RRID: AB_2799254), quinone oxidoreductase 1 (NQO-1; mouse, 1:2000, Cell Signaling Technology, Cat #3187, RRID: AB_2154354), and β-actin (mouse, 1:5000, CWBio, Beijing, China, Cat# CW0096, RRID: AB_2665433).

    Techniques: Expressing, shRNA, Immunofluorescence, Fluorescence, Comparison, Over Expression, Membrane

    Parkin increases PHB2/LC3-mediated mitophagy in SH-SY5Y cells. (A–D) Protein levels of Parkin, PHB2, and autophagy marker (LC3) in SH-SY5Y cells after treatment with Parkin-shRNA, Parkin-Over Exp, and MPP + (1 mM, 24 hours). (A) Protein expression levels of Parkin, LC3, and PHB2 in SH-SY5Y cells treated with Parkin-shRNA or Parkin-Over Exp under MPP + induction. (B–D) Quantitative analysis of Parkin, and LC3II/LC3I, PHB2 protein expression. (E) Immunofluorescence analysis of PHB2 and LC3. Cellular localization of LC3 and PHB2 in SH-SY5Y cells treated with Parkin-shRNA or Parkin-Over Exp under MPP + induction. Red fluorescence represents PHB2, green fluorescence represents LC3, and blue fluorescence represents DAPI. Original magnification 40×, Scale bar: 20 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs. control group; # P < 0.05,### P < 0.001, vs . MPP + group (one-way analysis of variance with Tukey's multiple comparison test). DAPI: 4′,6-Diamidino-2-phenylindole; LC3: microtubule-associated protein 1 light chain 3; MPP + : 1-methyl-4-phenylpyridinium; Over Exp: overexpression; Parkin: parkin RBR E3 ubiquitin-protein ligase; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line.

    Journal: Neural Regeneration Research

    Article Title: A novel mechanism of PHB2-mediated mitophagy participating in the development of Parkinson's disease

    doi: 10.4103/1673-5374.389356

    Figure Lengend Snippet: Parkin increases PHB2/LC3-mediated mitophagy in SH-SY5Y cells. (A–D) Protein levels of Parkin, PHB2, and autophagy marker (LC3) in SH-SY5Y cells after treatment with Parkin-shRNA, Parkin-Over Exp, and MPP + (1 mM, 24 hours). (A) Protein expression levels of Parkin, LC3, and PHB2 in SH-SY5Y cells treated with Parkin-shRNA or Parkin-Over Exp under MPP + induction. (B–D) Quantitative analysis of Parkin, and LC3II/LC3I, PHB2 protein expression. (E) Immunofluorescence analysis of PHB2 and LC3. Cellular localization of LC3 and PHB2 in SH-SY5Y cells treated with Parkin-shRNA or Parkin-Over Exp under MPP + induction. Red fluorescence represents PHB2, green fluorescence represents LC3, and blue fluorescence represents DAPI. Original magnification 40×, Scale bar: 20 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs. control group; # P < 0.05,### P < 0.001, vs . MPP + group (one-way analysis of variance with Tukey's multiple comparison test). DAPI: 4′,6-Diamidino-2-phenylindole; LC3: microtubule-associated protein 1 light chain 3; MPP + : 1-methyl-4-phenylpyridinium; Over Exp: overexpression; Parkin: parkin RBR E3 ubiquitin-protein ligase; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line.

    Article Snippet: Membranes were incubated overnight at 4°C in solutions with primary antibodies against PHB2 (rabbit, 1:2000, Proteintech, Chicago, IL, USA, Cat# 12295-1-AP, RRID: AB_2164779), Parkin (mouse, 1:2000, Abcam, Cambridge, UK, Cat# ab77924, RRID: AB_1566559), LC3B (rabbit, 1:2000, Abcam, Cat# ab192890, RRID: AB_2827794), p-PERK Thr980 (rabbit, 1:2000, Cell Signaling Technology, Danvers, MA, USA, Cat# 3179, RRID: AB_2095853), nuclear factor erythroid 2-related factor (Nrf2; rabbit, 1:2000, Cell Signaling Technology, Cat# 12721, RRID: AB_2715528), heme oxygenase 1 (HO-1; rabbit, 1:2000, Cell Signaling Technology, Cat# 43966, RRID: AB_2799254), quinone oxidoreductase 1 (NQO-1; mouse, 1:2000, Cell Signaling Technology, Cat #3187, RRID: AB_2154354), and β-actin (mouse, 1:5000, CWBio, Beijing, China, Cat# CW0096, RRID: AB_2665433).

    Techniques: Marker, shRNA, Expressing, Immunofluorescence, Fluorescence, Comparison, Over Expression

    Parkin regulates anti-oxidative stress protein expression via PHB2. (A–D) SH-SY5Y cells were treated with PHB2-shRNA, Parkin-Over Exp, and MPP + (1 mM, 24 hours). Changes in Nrf2, HO-1, and NQO-1 protein levels and quantitative analysis. (A) Protein expression levels of Nrf2, HO-1, and NQO-1 in SH-SY5Y cells treated with PHB2-shRNA or Parkin-Over Exp under MPP + induction. (B–D) Quantitative analysis of Nrf2, HO-1, and NQO-1 protein expression. Data are expressed as mean ± SEM ( n = 3). *** P < 0.001, vs. control group; # P < 0.05,### P < 0.001, vs. MPP + group (one-way analysis of variance with Tukey's multiple comparison test). HO-1: Heme oxygenase-1; MPP + : 1-methyl-4-phenylpyridinium; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; Over Exp: overexpression; Parkin: parkin RBR E3 ubiquitin-protein ligase; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line.

    Journal: Neural Regeneration Research

    Article Title: A novel mechanism of PHB2-mediated mitophagy participating in the development of Parkinson's disease

    doi: 10.4103/1673-5374.389356

    Figure Lengend Snippet: Parkin regulates anti-oxidative stress protein expression via PHB2. (A–D) SH-SY5Y cells were treated with PHB2-shRNA, Parkin-Over Exp, and MPP + (1 mM, 24 hours). Changes in Nrf2, HO-1, and NQO-1 protein levels and quantitative analysis. (A) Protein expression levels of Nrf2, HO-1, and NQO-1 in SH-SY5Y cells treated with PHB2-shRNA or Parkin-Over Exp under MPP + induction. (B–D) Quantitative analysis of Nrf2, HO-1, and NQO-1 protein expression. Data are expressed as mean ± SEM ( n = 3). *** P < 0.001, vs. control group; # P < 0.05,### P < 0.001, vs. MPP + group (one-way analysis of variance with Tukey's multiple comparison test). HO-1: Heme oxygenase-1; MPP + : 1-methyl-4-phenylpyridinium; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; Over Exp: overexpression; Parkin: parkin RBR E3 ubiquitin-protein ligase; PHB2: prohibitin 2; SH-SY5Y: human neuroblastoma cell line.

    Article Snippet: Membranes were incubated overnight at 4°C in solutions with primary antibodies against PHB2 (rabbit, 1:2000, Proteintech, Chicago, IL, USA, Cat# 12295-1-AP, RRID: AB_2164779), Parkin (mouse, 1:2000, Abcam, Cambridge, UK, Cat# ab77924, RRID: AB_1566559), LC3B (rabbit, 1:2000, Abcam, Cat# ab192890, RRID: AB_2827794), p-PERK Thr980 (rabbit, 1:2000, Cell Signaling Technology, Danvers, MA, USA, Cat# 3179, RRID: AB_2095853), nuclear factor erythroid 2-related factor (Nrf2; rabbit, 1:2000, Cell Signaling Technology, Cat# 12721, RRID: AB_2715528), heme oxygenase 1 (HO-1; rabbit, 1:2000, Cell Signaling Technology, Cat# 43966, RRID: AB_2799254), quinone oxidoreductase 1 (NQO-1; mouse, 1:2000, Cell Signaling Technology, Cat #3187, RRID: AB_2154354), and β-actin (mouse, 1:5000, CWBio, Beijing, China, Cat# CW0096, RRID: AB_2665433).

    Techniques: Expressing, shRNA, Comparison, Over Expression

    Silencing PHB2 inhibits mitophagy and aggravates dopaminergic neuronal loss in PD mice. (A) Schematic of PHB2-shRNA and MPTP treatment in mice. (B, C) Quantitative analysis of PHB2 protein levels in the midbrain of C57BL/6J mice after PHB2-shRNA injection. (B) Protein expression levels of PHB2 in C57BL/6J mice treated with PHB2-shRNA. (C) Quantitative analysis of PHB2 protein expression. (D–F) An acute PD model was established by intraperitoneal injection of MPTP in C57BL/6J mice after PHB2-shRNA injection. Changes and quantitative analysis of LC3II/LC3I and PHB2 protein levels in the midbrain. (D) Protein expression levels of PHB2 and LC3 after MPTP injection in PD model mice. (E, F) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (G, H) TH immunofluorescence (Alexa Fluor 488, green fluorescence) shows dopaminergic neurons in the substantia nigra in PD mice with silenced PHB2. (G) Immunofluorescence of TH-positive neurons in the substantia nigra. Mice were treated with PHB2-shRNA and MPTP. Original magnification 10×, Scale bar: 200 μm. (H) Quantitative analysis of TH-positive neurons in the substantia nigra. (I) Fluorescence co-localization of LC3 and PHB2, TIM23, or TOM20 in the substantia nigra. Mice were treated with PHB2-shRNA and MPTP. Red fluorescence (Alexa Fluor 555): PHB2, TOM20, and TIM23, green fluorescence (Alexa Fluor 488): LC3, and blue fluorescence: DAPI. Original magnification 20×, Scale bars: 20 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, *** P < 0.001, vs. control group; $ P < 0.05, $$ P < 0.01, $$$ P < 0.001, vs. MPTP group (one-way analysis of variance with Tukey's multiple comparison test). LC3: Microtubule-associated protein 1 light chain 3; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; PD: Parkinson's disease; PHB2: Prohibitin 2; TH: tyrosine hydroxylase; TIM23: translocase of inner mitochondrial membrane 23; TOM20: translocase of outer mitochondrial membrane 20.

    Journal: Neural Regeneration Research

    Article Title: A novel mechanism of PHB2-mediated mitophagy participating in the development of Parkinson's disease

    doi: 10.4103/1673-5374.389356

    Figure Lengend Snippet: Silencing PHB2 inhibits mitophagy and aggravates dopaminergic neuronal loss in PD mice. (A) Schematic of PHB2-shRNA and MPTP treatment in mice. (B, C) Quantitative analysis of PHB2 protein levels in the midbrain of C57BL/6J mice after PHB2-shRNA injection. (B) Protein expression levels of PHB2 in C57BL/6J mice treated with PHB2-shRNA. (C) Quantitative analysis of PHB2 protein expression. (D–F) An acute PD model was established by intraperitoneal injection of MPTP in C57BL/6J mice after PHB2-shRNA injection. Changes and quantitative analysis of LC3II/LC3I and PHB2 protein levels in the midbrain. (D) Protein expression levels of PHB2 and LC3 after MPTP injection in PD model mice. (E, F) Quantitative analysis of PHB2 and LC3II/LC3I protein expression. (G, H) TH immunofluorescence (Alexa Fluor 488, green fluorescence) shows dopaminergic neurons in the substantia nigra in PD mice with silenced PHB2. (G) Immunofluorescence of TH-positive neurons in the substantia nigra. Mice were treated with PHB2-shRNA and MPTP. Original magnification 10×, Scale bar: 200 μm. (H) Quantitative analysis of TH-positive neurons in the substantia nigra. (I) Fluorescence co-localization of LC3 and PHB2, TIM23, or TOM20 in the substantia nigra. Mice were treated with PHB2-shRNA and MPTP. Red fluorescence (Alexa Fluor 555): PHB2, TOM20, and TIM23, green fluorescence (Alexa Fluor 488): LC3, and blue fluorescence: DAPI. Original magnification 20×, Scale bars: 20 μm. Data are expressed as mean ± SEM ( n = 3). * P < 0.05, *** P < 0.001, vs. control group; $ P < 0.05, $$ P < 0.01, $$$ P < 0.001, vs. MPTP group (one-way analysis of variance with Tukey's multiple comparison test). LC3: Microtubule-associated protein 1 light chain 3; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; PD: Parkinson's disease; PHB2: Prohibitin 2; TH: tyrosine hydroxylase; TIM23: translocase of inner mitochondrial membrane 23; TOM20: translocase of outer mitochondrial membrane 20.

    Article Snippet: Membranes were incubated overnight at 4°C in solutions with primary antibodies against PHB2 (rabbit, 1:2000, Proteintech, Chicago, IL, USA, Cat# 12295-1-AP, RRID: AB_2164779), Parkin (mouse, 1:2000, Abcam, Cambridge, UK, Cat# ab77924, RRID: AB_1566559), LC3B (rabbit, 1:2000, Abcam, Cat# ab192890, RRID: AB_2827794), p-PERK Thr980 (rabbit, 1:2000, Cell Signaling Technology, Danvers, MA, USA, Cat# 3179, RRID: AB_2095853), nuclear factor erythroid 2-related factor (Nrf2; rabbit, 1:2000, Cell Signaling Technology, Cat# 12721, RRID: AB_2715528), heme oxygenase 1 (HO-1; rabbit, 1:2000, Cell Signaling Technology, Cat# 43966, RRID: AB_2799254), quinone oxidoreductase 1 (NQO-1; mouse, 1:2000, Cell Signaling Technology, Cat #3187, RRID: AB_2154354), and β-actin (mouse, 1:5000, CWBio, Beijing, China, Cat# CW0096, RRID: AB_2665433).

    Techniques: shRNA, Injection, Expressing, Immunofluorescence, Fluorescence, Comparison, Membrane

    Silencing PHB2 reduces antioxidative stress protein expression and aggravates motor defecits in PD mice. (A–D) Changes and quantitative analysis of Nrf2, HO-1, and NQO-1 protein levels in the midbrain of PHB2-shRNA and MPTP-treated PD mice. (E) Quantitative analysis of mice in the tail suspension and rotarod tests. Data are expressed as mean ± SEM ( n = 5). * P < 0.05, ** P < 0.01, vs. control group; $ P < 0.05, $$$ P < 0.001, vs . MPTP group (one-way analysis of variance with Tukey's multiple comparison test). HO-1: Heme oxygenase-1; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; PD: Parkinson's disease; PHB2: prohibitin 2.

    Journal: Neural Regeneration Research

    Article Title: A novel mechanism of PHB2-mediated mitophagy participating in the development of Parkinson's disease

    doi: 10.4103/1673-5374.389356

    Figure Lengend Snippet: Silencing PHB2 reduces antioxidative stress protein expression and aggravates motor defecits in PD mice. (A–D) Changes and quantitative analysis of Nrf2, HO-1, and NQO-1 protein levels in the midbrain of PHB2-shRNA and MPTP-treated PD mice. (E) Quantitative analysis of mice in the tail suspension and rotarod tests. Data are expressed as mean ± SEM ( n = 5). * P < 0.05, ** P < 0.01, vs. control group; $ P < 0.05, $$$ P < 0.001, vs . MPTP group (one-way analysis of variance with Tukey's multiple comparison test). HO-1: Heme oxygenase-1; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; NQO-1: NAD(P)H quinone dehydrogenase 1; Nrf2: nuclear factor erythroid 2-related factor 2; PD: Parkinson's disease; PHB2: prohibitin 2.

    Article Snippet: Membranes were incubated overnight at 4°C in solutions with primary antibodies against PHB2 (rabbit, 1:2000, Proteintech, Chicago, IL, USA, Cat# 12295-1-AP, RRID: AB_2164779), Parkin (mouse, 1:2000, Abcam, Cambridge, UK, Cat# ab77924, RRID: AB_1566559), LC3B (rabbit, 1:2000, Abcam, Cat# ab192890, RRID: AB_2827794), p-PERK Thr980 (rabbit, 1:2000, Cell Signaling Technology, Danvers, MA, USA, Cat# 3179, RRID: AB_2095853), nuclear factor erythroid 2-related factor (Nrf2; rabbit, 1:2000, Cell Signaling Technology, Cat# 12721, RRID: AB_2715528), heme oxygenase 1 (HO-1; rabbit, 1:2000, Cell Signaling Technology, Cat# 43966, RRID: AB_2799254), quinone oxidoreductase 1 (NQO-1; mouse, 1:2000, Cell Signaling Technology, Cat #3187, RRID: AB_2154354), and β-actin (mouse, 1:5000, CWBio, Beijing, China, Cat# CW0096, RRID: AB_2665433).

    Techniques: Expressing, shRNA, Suspension, Comparison

    Journal: Molecular Cell

    Article Title: Identification of TMEM126A as OXA1L-interacting protein reveals cotranslational quality control in mitochondria

    doi: 10.1016/j.molcel.2023.12.013

    Figure Lengend Snippet:

    Article Snippet: Rabbit polyclonal anti-PHB2 , Proteintech , Cat#12295-1-AP; RRID: AB_2164779.

    Techniques: Recombinant, Activity Assay, Software

    APOL1- and APOL3-specific interactions (A) Interaction of APOL1, APOL1 mutants, or APOL3 with PHB2, as measured in the E. coli pDUET co-expression system. Nickel binding of S-tagged proteins (yellow frame) reflects their relative association with His-tagged partners (green frame). These data are representative of at least 3 independent experiments. A1, WT APOL1; Δ, APOL1Δ; A3, WT APOL3; H4Q, APOL1 mutHel4Q; H5Q, APOL1 mutHel5Q; H5P, APOL1 mutHel5P; H5mS, APOL1 mutHel5S; H5sw, APOL1 Hel5 swap; BH3E, APOL1 BH3 mutant E. (B) Representative SPR sensorgrams of immobilized L1Hel5 (left) or L3Hel5 (right) peptide interaction with recombinant PHB2 with or without calcium (n = 3). (C) Representative SPR sensorgrams of immobilized L1Hel5 (left) or L3Hel5 (right) peptide interaction with APOL3 Hel5, NTD, or CTD peptide (n = 3). (D) SPR measurements of bound sVAMP8 (left) or TbVAMP7Bp (right) interaction with various APOL3 versions as indicated (n = 3). (E) SPR measurements of bound sVAMP8 (left) or TbVAMP7Bp (right) interaction with different APOL3 peptides as indicated (n = 3). (F) Immunodetection of recombinant sVAMP8 binding to various lipids spotted on membrane strips. LPA, lysophosphatidic acid; LPC, lysophosphocholine; PI, phosphatidylinositol; PE, phosphatidylethanolamine; PC, phosphatidylcholine; S1P, sphingosine-1-phosphate; PA, phosphatidic acid; PS, phosphatidylserine; TG, triglyceride; DAG, diacylglycerol; PG, phosphatidylglycerol; CL, cardiolipin; Chol, cholesterol; SM, sphingomyelin. Blank, no lipid.

    Journal: Cell Reports

    Article Title: Apolipoproteins L1 and L3 control mitochondrial membrane dynamics

    doi: 10.1016/j.celrep.2023.113528

    Figure Lengend Snippet: APOL1- and APOL3-specific interactions (A) Interaction of APOL1, APOL1 mutants, or APOL3 with PHB2, as measured in the E. coli pDUET co-expression system. Nickel binding of S-tagged proteins (yellow frame) reflects their relative association with His-tagged partners (green frame). These data are representative of at least 3 independent experiments. A1, WT APOL1; Δ, APOL1Δ; A3, WT APOL3; H4Q, APOL1 mutHel4Q; H5Q, APOL1 mutHel5Q; H5P, APOL1 mutHel5P; H5mS, APOL1 mutHel5S; H5sw, APOL1 Hel5 swap; BH3E, APOL1 BH3 mutant E. (B) Representative SPR sensorgrams of immobilized L1Hel5 (left) or L3Hel5 (right) peptide interaction with recombinant PHB2 with or without calcium (n = 3). (C) Representative SPR sensorgrams of immobilized L1Hel5 (left) or L3Hel5 (right) peptide interaction with APOL3 Hel5, NTD, or CTD peptide (n = 3). (D) SPR measurements of bound sVAMP8 (left) or TbVAMP7Bp (right) interaction with various APOL3 versions as indicated (n = 3). (E) SPR measurements of bound sVAMP8 (left) or TbVAMP7Bp (right) interaction with different APOL3 peptides as indicated (n = 3). (F) Immunodetection of recombinant sVAMP8 binding to various lipids spotted on membrane strips. LPA, lysophosphatidic acid; LPC, lysophosphocholine; PI, phosphatidylinositol; PE, phosphatidylethanolamine; PC, phosphatidylcholine; S1P, sphingosine-1-phosphate; PA, phosphatidic acid; PS, phosphatidylserine; TG, triglyceride; DAG, diacylglycerol; PG, phosphatidylglycerol; CL, cardiolipin; Chol, cholesterol; SM, sphingomyelin. Blank, no lipid.

    Article Snippet: Rabbit anti-PHB2 , Proteintech , Cat# 12295-1-AP; RRID: AB_2164779.

    Techniques: Expressing, Binding Assay, Mutagenesis, Recombinant, Immunodetection, Membrane

    Journal: Cell Reports

    Article Title: Apolipoproteins L1 and L3 control mitochondrial membrane dynamics

    doi: 10.1016/j.celrep.2023.113528

    Figure Lengend Snippet:

    Article Snippet: Rabbit anti-PHB2 , Proteintech , Cat# 12295-1-AP; RRID: AB_2164779.

    Techniques: Recombinant, Kinase Assay, Stripping Membranes, Software