Journal: Clinical and Translational Medicine

Article Title: PINK1 modulates Prdx2 to reduce lipotoxicity‐induced apoptosis and attenuate cardiac dysfunction in heart failure mice with a preserved ejection fraction

doi: 10.1002/ctm2.70166

Figure Lengend Snippet: PTEN‐induced kinase 1 (PINK1) deficiency reduces peroxiredoxin‐2 (Prdx2) and exacerbates JNK and p38 phosphorylation leading to myocardial apoptosis. (A) Western blotting for the expression of PINK1, Prdx2, phospho‐p38, p38, phospho‐JNK, JNK, Bcl2, Bax, Cleaved‐Caspase3 and β‐actin in heart tissues. (B) Quantitative analyses of protein expression of PINK1, Prdx2, phospho‐p38, p38, phospho‐JNK, JNK, Bcl2, Bax, Cleaved‐Caspase3 and β‐actin. (C) immunohistochemical of PINK1 in heart tissues of WT and PINK1‐KO mice. Scale bar, 50 µm. (D) immunohistochemical of Prdx2 in heart tissues of WT and PINK1‐KO mice. Scale bar, 50 µm. (E) The apoptosis rate of cardiomyocytes by TUNEL assay Scale bar, 100 µm. For all statistical plots, the data are presented as the means ± SEs. * p < 0.05; ** p < 0.01; *** p < 0.001 using Student t test.

Article Snippet: The antibodies used in this study included anti‐PINK1 (Santa Cruz, sc‐517353, rabbit) and the following from Proteintech: anti‐Prdx2 (10545‐2‐AP, rabbit), anti‐P‐JNK (80024‐1‐RR, rabbit), anti‐P‐p38 (28796‐1‐AP, rabbit), anti‐JNK (24164‐1‐AP, rabbit), anti‐p38 (14064‐1‐AP, rabbit), anti‐Bcl2 (26593‐1‐AP, rabbit), anti‐Bax (50599‐2‐Ig, rabbit), anti‐C‐Caspase3 (68773‐1‐Ig, mouse), anti‐GFP (50430‐2‐AP, rabbit), anti‐ANP (27426‐1‐AP, rabbit) and anti‐β‐actin (66009‐1‐Ig, mouse).

Techniques: Western Blot, Expressing, Immunohistochemical staining, TUNEL Assay

Journal: Clinical and Translational Medicine

Article Title: PINK1 modulates Prdx2 to reduce lipotoxicity‐induced apoptosis and attenuate cardiac dysfunction in heart failure mice with a preserved ejection fraction

doi: 10.1002/ctm2.70166

Figure Lengend Snippet: PTEN‐induced kinase 1 (PINK1) overexpression attenuates myocardial apoptosis by restoring peroxiredoxin‐2 (Prdx2). (A) Western blotting for the expression of PINK1, Prdx2, phospho‐p38, p38, phospho‐JNK, JNK, Bcl2, Bax, Cleaved‐Caspase3 and β‐actin in heart tissues. (B) Quantitative analyses of protein expression of PINK1, Prdx2, phospho‐p38, p38, phospho‐JNK, JNK, Bcl2, Bax, Cleaved‐Caspase3 and β‐actin. (C) immunohistochemical of PINK1 in heart tissues of WT and PINK1‐KO mice. Scale bar, 50 µm. (D) immunohistochemical of Prdx2 in heart tissues of WT and PINK1‐KO mice. Scale bar, 50 µm. (E) The apoptosis rate of cardiomyocytes by TUNEL assay Scale bar, 100 µm. For all statistical plots, the data are presented as the means ± SEs. * p < 0.05; ** p < 0.01; *** p < 0.001 using Student t test.

Article Snippet: The antibodies used in this study included anti‐PINK1 (Santa Cruz, sc‐517353, rabbit) and the following from Proteintech: anti‐Prdx2 (10545‐2‐AP, rabbit), anti‐P‐JNK (80024‐1‐RR, rabbit), anti‐P‐p38 (28796‐1‐AP, rabbit), anti‐JNK (24164‐1‐AP, rabbit), anti‐p38 (14064‐1‐AP, rabbit), anti‐Bcl2 (26593‐1‐AP, rabbit), anti‐Bax (50599‐2‐Ig, rabbit), anti‐C‐Caspase3 (68773‐1‐Ig, mouse), anti‐GFP (50430‐2‐AP, rabbit), anti‐ANP (27426‐1‐AP, rabbit) and anti‐β‐actin (66009‐1‐Ig, mouse).

Techniques: Over Expression, Western Blot, Expressing, Immunohistochemical staining, TUNEL Assay

Journal: Clinical and Translational Medicine

Article Title: PINK1 modulates Prdx2 to reduce lipotoxicity‐induced apoptosis and attenuate cardiac dysfunction in heart failure mice with a preserved ejection fraction

doi: 10.1002/ctm2.70166

Figure Lengend Snippet: PTEN‐induced kinase 1 (PINK1) interacts with peroxiredoxin‐2 (Prdx2) and both are impaired in lipotoxicity. (A) Immunoprecipitation of PINK1 with Prdx2 in NRCM. (B) two alpha structure of PINK1: N‐terminal region (amino acids 1133) and C‐terminal region (amino acids 138500). (C) Detection of immunoprecipitation protein bands in a gel by silver staining. (D) Immunoblot of immunoprecipitation for the GFP and Prdx2. (E) Immunofluorescence co‐localisation of PINK1 with Prdx2 in NRCM. Scale bar, 25 µm. (F) Detection of LDs by Fluorescent Microscopy in palmitic acid gradient concentration. Scale bar, 25 µm. (G) Quantitative analyses of LDs. (H) Western blotting for the expression of PINK1, Prdx2, ANP and β‐actin in palmitic acid gradient concentration. (I) Cell viability of NRCM in palmitic acid gradient concentration. For all statistical plots, the data are presented as the means ± SEs. * p < 0.05; ** p < 0.01; *** p < 0.001 using Student t test.

Article Snippet: The antibodies used in this study included anti‐PINK1 (Santa Cruz, sc‐517353, rabbit) and the following from Proteintech: anti‐Prdx2 (10545‐2‐AP, rabbit), anti‐P‐JNK (80024‐1‐RR, rabbit), anti‐P‐p38 (28796‐1‐AP, rabbit), anti‐JNK (24164‐1‐AP, rabbit), anti‐p38 (14064‐1‐AP, rabbit), anti‐Bcl2 (26593‐1‐AP, rabbit), anti‐Bax (50599‐2‐Ig, rabbit), anti‐C‐Caspase3 (68773‐1‐Ig, mouse), anti‐GFP (50430‐2‐AP, rabbit), anti‐ANP (27426‐1‐AP, rabbit) and anti‐β‐actin (66009‐1‐Ig, mouse).

Techniques: Immunoprecipitation, Silver Staining, Western Blot, Immunofluorescence, Microscopy, Concentration Assay, Expressing

Journal: Clinical and Translational Medicine

Article Title: PINK1 modulates Prdx2 to reduce lipotoxicity‐induced apoptosis and attenuate cardiac dysfunction in heart failure mice with a preserved ejection fraction

doi: 10.1002/ctm2.70166

Figure Lengend Snippet: siPINK1 aggravates neonatal rat cardiomyocyte (NRCM) lipotoxicity through peroxiredoxin‐2 (Prdx2) reduction. (A) Western blotting for the expression of PINK1, Prdx2, phospho‐p38, p38, phospho‐JNK, JNK, Bcl2, Bax, Cleaved‐Caspase3 and β‐actin in myocardial lipotoxicity. (B) immunofluorescence of PINK1 in myocardial lipotoxicity. (C) Quantitative analyses of PINK1 positive area. (D) immunofluorescence of Prdx2 in myocardial lipotoxicity. (E) DCHF staining to reflect myocardial ROS production. Scale bar, 50 µm. For all statistical plots, the data are presented as the means ± SEs. * p < 0.05; ** p < 0.01; *** p < 0.001 using Student t test.

Article Snippet: The antibodies used in this study included anti‐PINK1 (Santa Cruz, sc‐517353, rabbit) and the following from Proteintech: anti‐Prdx2 (10545‐2‐AP, rabbit), anti‐P‐JNK (80024‐1‐RR, rabbit), anti‐P‐p38 (28796‐1‐AP, rabbit), anti‐JNK (24164‐1‐AP, rabbit), anti‐p38 (14064‐1‐AP, rabbit), anti‐Bcl2 (26593‐1‐AP, rabbit), anti‐Bax (50599‐2‐Ig, rabbit), anti‐C‐Caspase3 (68773‐1‐Ig, mouse), anti‐GFP (50430‐2‐AP, rabbit), anti‐ANP (27426‐1‐AP, rabbit) and anti‐β‐actin (66009‐1‐Ig, mouse).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining

Journal: Clinical and Translational Medicine

Article Title: PINK1 modulates Prdx2 to reduce lipotoxicity‐induced apoptosis and attenuate cardiac dysfunction in heart failure mice with a preserved ejection fraction

doi: 10.1002/ctm2.70166

Figure Lengend Snippet: adPINK1 attenuates neonatal rat cardiomyocyte (NRCM) lipotoxicity by increasing peroxiredoxin‐2 (Prdx2). (A) Western blotting for the expression of PINK1, Prdx2, phospho‐p38, p38, phospho‐JNK, JNK, Bcl2, Bax, Cleaved‐Caspase3 and β‐actin in myocardial lipotoxicity. (B) immunofluorescence of PINK1 in myocardial lipotoxicity. (C) Quantitative analyses of PINK1 positive area. (D) immunofluorescence of Prdx2 in myocardial lipotoxicity. (E) DCHF staining to reflect myocardial ROS production. Scale bar, 50 µm. For all statistical plots, the data are presented as the means ± SEs. * p < 0.05; ** p < 0.01; *** p < 0.001 using Student t test.

Article Snippet: The antibodies used in this study included anti‐PINK1 (Santa Cruz, sc‐517353, rabbit) and the following from Proteintech: anti‐Prdx2 (10545‐2‐AP, rabbit), anti‐P‐JNK (80024‐1‐RR, rabbit), anti‐P‐p38 (28796‐1‐AP, rabbit), anti‐JNK (24164‐1‐AP, rabbit), anti‐p38 (14064‐1‐AP, rabbit), anti‐Bcl2 (26593‐1‐AP, rabbit), anti‐Bax (50599‐2‐Ig, rabbit), anti‐C‐Caspase3 (68773‐1‐Ig, mouse), anti‐GFP (50430‐2‐AP, rabbit), anti‐ANP (27426‐1‐AP, rabbit) and anti‐β‐actin (66009‐1‐Ig, mouse).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining

Journal: Clinical and Translational Medicine

Article Title: PINK1 modulates Prdx2 to reduce lipotoxicity‐induced apoptosis and attenuate cardiac dysfunction in heart failure mice with a preserved ejection fraction

doi: 10.1002/ctm2.70166

Figure Lengend Snippet: siPrdx2 attenuates the alleviating effect of adPINK1 on neonatal rat cardiomyocyte (NRCM) lipotoxicity. (A) Western blotting for the expression of PINK1, Prdx2, phospho‐p38, p38, phospho‐JNK, JNK, Bcl2, Bax, Cleaved‐Caspase3 and β‐actin in myocardial lipotoxicity. (B) Quantitative analyses of protein expression of PINK1, Prdx2, phospho‐p38, p38, phospho‐JNK, JNK, Bcl2, Bax, Cleaved‐Caspase3 and β‐actin. (C) immunofluorescence of PINK1 in myocardial lipotoxicity. (D) Quantitative analyses of PINK1 positive area. (E) immunofluorescence of Prdx2 in myocardial lipotoxicity. (F) Quantitative analyses of Prdx2 positive area. (G) DCHF staining to reflect myocardial ROS production. Scale bar, 50 µm. (H) Quantitative analyses of DCFH staining. (I) The apoptosis rate of cardiomyocytes by TUNEL assay. Scale bar, 100 µm. (J) Quantitative analyses of TUNEL positive area. For all statistical plots, the data are presented as the means ± SEs. * p < 0.05; ** p < 0.01; *** p < 0.001 using Student t test.

Article Snippet: The antibodies used in this study included anti‐PINK1 (Santa Cruz, sc‐517353, rabbit) and the following from Proteintech: anti‐Prdx2 (10545‐2‐AP, rabbit), anti‐P‐JNK (80024‐1‐RR, rabbit), anti‐P‐p38 (28796‐1‐AP, rabbit), anti‐JNK (24164‐1‐AP, rabbit), anti‐p38 (14064‐1‐AP, rabbit), anti‐Bcl2 (26593‐1‐AP, rabbit), anti‐Bax (50599‐2‐Ig, rabbit), anti‐C‐Caspase3 (68773‐1‐Ig, mouse), anti‐GFP (50430‐2‐AP, rabbit), anti‐ANP (27426‐1‐AP, rabbit) and anti‐β‐actin (66009‐1‐Ig, mouse).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, TUNEL Assay

Journal: Redox biology

Article Title: Cytoglobin scavenges intracellular hydrogen peroxide and regulates redox signals in the vasculature.

doi: 10.1016/j.redox.2025.103633

Figure Lengend Snippet: Fig. 4. Cytoglobin decreases peroxiredoxin hyperoxidation and maintains active peroxiredoxin 2. a, Control (Empty Vector) and cytoglobin (CYGB) expressing HEK293 cells were exposed to hydrogen peroxide for 10 min followed by the addition of NEM. Lysates were used for non-reducing SDS PAGE and Western blotting to examine protein levels of hyperoxidized peroxiredoxins (Prx-SO2/3) using beta actin (ACTB) as an internal reference. Cytoglobin levels (CYGB) are also shown. Right panel, quantitation of experiments described in left panel. b and c, Control (Empty Vector) and cytoglobin (CYGB) expressing HEK293 cells were exposed to 150 μM hydrogen peroxide for various amounts of time followed by the addition of NEM. Lysates were used in reducing and non-reducing SDS PAGE and Western blotting to examine protein levels of peroxiredoxin 1 (b, PRXD1) and 2 (c, PRXD2). Arrows indicate the position of the dimers and monomers. Right panels, quantitation of experiments represented in left panels. All graphs show mean ± SEM and each point represents one independent experimental replicate. Statistical analysis was performed using two-way ANOVA with Tukey’s multicomparisons test.

Article Snippet: Blots were incubated with primary antibodies for PRX-SO2/3 (1:1500), PRDX1(1:10 000), PRDX2 (1:10 000) or CYGB (1:2000) for 1 h at room temperature, washed 3 × 15 min, incubated with either goat anti mouse or goat anti rabbit-HRP secondary antibodies (1:2500) for 1 h at RT, washed 3 × 15 min, then visualized using BioRad Clarity Western ECL substrate on a BioRad ChemiDoc MP imaging system.

Techniques: Control, Plasmid Preparation, Expressing, SDS Page, Western Blot, Quantitation Assay

Journal: Redox biology

Article Title: Cytoglobin scavenges intracellular hydrogen peroxide and regulates redox signals in the vasculature.

doi: 10.1016/j.redox.2025.103633

Figure Lengend Snippet: Fig. 5. The peroxidase activity of cytoglobin is competitive with peroxiredoxin 2 hyperoxidation in vitro. a, Non-reducing SDS-PAGE gels of reduced peroxiredoxin 2 (2.5 μM) incubated with hydrogen peroxide (H2O2, 25 μM) for 5 min in the presence of increasing concentrations of cytoglobin. Representative of three independent experiments. b, Time-dependent decomposition of hydrogen peroxide (25 μM) by cytoglobin (CYGB; 2.5 μM) or peroxiredoxin 2 (PRDX2; 2.5 μM) with DTPA (100 μM) and 100 mM phosphate buffer, in the presence or absence of DTT (2 mM). The reduction of hydrogen peroxide in the presence of DTT alone is also shown. c, Effect of potassium cyanide (KCN; 100 μM) on the decomposition of hydrogen peroxide (25 μM) by cytoglobin (1.25 and 2.5 μM). d, Peroxidase activity of WT, C38G, and C83G cytoglobin (1.25 μM each) with 25 μM hydrogen peroxide. Results are means of triplicates.

Article Snippet: Blots were incubated with primary antibodies for PRX-SO2/3 (1:1500), PRDX1(1:10 000), PRDX2 (1:10 000) or CYGB (1:2000) for 1 h at room temperature, washed 3 × 15 min, incubated with either goat anti mouse or goat anti rabbit-HRP secondary antibodies (1:2500) for 1 h at RT, washed 3 × 15 min, then visualized using BioRad Clarity Western ECL substrate on a BioRad ChemiDoc MP imaging system.

Techniques: Activity Assay, In Vitro, SDS Page, Incubation

Journal: PLoS ONE

Article Title: Transcript and Protein Analysis Reveals Better Survival Skills of Monocyte-Derived Dendritic Cells Compared to Monocytes during Oxidative Stress

doi: 10.1371/journal.pone.0043357

Figure Lengend Snippet: PCR profiling array analysis of oxidative stress related genes (OSRG) with two-fold or more up-or downregulated expression during differentiation of monocytes(mono) into monocyte-derived DCs (moDC).

Article Snippet: Homo sapiens-specific Taqman gene expression assays (Applied biosystems, California, USA) were purchased for dual oxidase (DUOX)-1 (Hs00213694_m1), superoxide dismutase 2 (SOD2, Hs00167309_m1), glutathione peroxidise-3 (GPX3) (Hs00173566_m1), GPX7 (Hs00210410_m1), PRDX2 (Hs03044902_g1), as well as for an endogenous control ACTB (Hs99999903_m1).

Techniques: Expressing, Clinical Proteomics

Journal: PLoS ONE

Article Title: Transcript and Protein Analysis Reveals Better Survival Skills of Monocyte-Derived Dendritic Cells Compared to Monocytes during Oxidative Stress

doi: 10.1371/journal.pone.0043357

Figure Lengend Snippet: Relative mRNA expression of DUOX1, PRDX2, GPX3, SOD2 and GPX7 in monocytes and moDCs . mRNA expression was analyzed using Taqman assays in individual non-pooled RNA extracts. Data were normalized to expression of ACTB and monocytes served as control group (expression set to 1). B. Western blot analysis of DUOX1, PRDX2, SOD2 and GPX7 during differentiation from monocytes (M) to moDCs (DC). The antibody against GPX3 did not yield signals that were distinguishable from background noise in the immunoblots. Representative blots (top panels) and mean densitometric ratios of protein of interest/ACTB. Values show mean ± SEM, n = 4. N.D. = not detectable. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Homo sapiens-specific Taqman gene expression assays (Applied biosystems, California, USA) were purchased for dual oxidase (DUOX)-1 (Hs00213694_m1), superoxide dismutase 2 (SOD2, Hs00167309_m1), glutathione peroxidise-3 (GPX3) (Hs00173566_m1), GPX7 (Hs00210410_m1), PRDX2 (Hs03044902_g1), as well as for an endogenous control ACTB (Hs99999903_m1).

Techniques: Expressing, Control, Western Blot

Journal: PLoS ONE

Article Title: Transcript and Protein Analysis Reveals Better Survival Skills of Monocyte-Derived Dendritic Cells Compared to Monocytes during Oxidative Stress

doi: 10.1371/journal.pone.0043357

Figure Lengend Snippet: A. Reduced PRDX2 mRNA levels in unstimulated moDCs after transfection with anti-PRDX2 siRNA [qPCR; 2 −ΔΔCt normalization using ACTB as reference gene and the non-transfected sample (---) as control (set to 1, n = 4)]. B. Reduction of PRDX2 protein in unstimulated moDCs monitored by Western Blot. Data were normalized using ACTB as reference gene (n = 4). C. CM-DCF fluorescence was used as index for intracellular ROS levels in non-transfected moDCs and moDCs transfected with PRDX2-targeting siRNA, with or without tert-BHP stimulation (200 μM, 30 min, n = 8). D. Neutral Red assay was used as index viability index of non-transfected moDCs and moDCs transfected with PRDX2-targeting siRNA, with or without tert-BHP stimulation (200 μM, 30 min, n = 8). **P<0.01 vs. untransfected (--) and negative control (neg. ctr), xxx P<0.001 vs. Unstimulated, paired Student's t-test (two-tailed).

Article Snippet: Homo sapiens-specific Taqman gene expression assays (Applied biosystems, California, USA) were purchased for dual oxidase (DUOX)-1 (Hs00213694_m1), superoxide dismutase 2 (SOD2, Hs00167309_m1), glutathione peroxidise-3 (GPX3) (Hs00173566_m1), GPX7 (Hs00210410_m1), PRDX2 (Hs03044902_g1), as well as for an endogenous control ACTB (Hs99999903_m1).

Techniques: Transfection, Control, Western Blot, Fluorescence, Neutral Red Assay, Negative Control, Two Tailed Test

Journal: The American journal of pathology

Article Title: Low-dose-rate, low-dose irradiation delays neurodegeneration in a model of retinitis pigmentosa.

doi: 10.1016/j.ajpath.2011.09.025

Figure Lengend Snippet: Figure 4. Prdx2 plays a major role in the neuroprotective effects induced by low-dose-rate, low-dose (LDR-LD) radiation. A: Time course of Prdx2 mRNA levels in LDR-LD–irradiated retinas (n 6). B: Western blot analysis of Prdx2 in LDR-LD–irradiated retinas (n 9). C–F: In situ hybridization study of Prdx2 in the retina. G–J: Immunohistochemistry of Prdx2 in the retina. K: Relative expression level of Prdx2 mRNA 26 hours after intravitreal siRNA injection. Radiation was administered at 24 hours after injection. NC, negative control injected; Rad, LDR-LD radiation (n 12). A normal C57BL/6 strain was used in experiments A to K. L: PRL thickness of an rd10 (P25) mouse retina. Intravitreal injection of siRNA at P18. LDR-LD radiation was performed 24 hours after the siRNA injection (P19) (n 12). Scale bars: 60 m (C–J). *P 0.05, **P 0.01 versus control. P values were from post hoc analysis.

Article Snippet: For the Prdx2 blot, a rabbit polyclonal antibody to Prdx2 (ProteinTech Group, Chicago, IL) was used followed by goat anti-rabbit IgG antibody conjugated to horseradish peroxidase (Vector Laboratories, Burlingame, CA).

Techniques: Irradiation, Western Blot, In Situ Hybridization, Immunohistochemistry, Expressing, Injection, Negative Control, Control

Journal: Physiological Reports

Article Title: Active hematopoiesis triggers exosomal release of PRDX2 that promotes osteoclast formation

doi: 10.14814/phy2.14745

Figure Lengend Snippet: Identification of L‐plastin and PRDXs in K562 cells. Conditioned media (CM) was collected and TCA precipitated, and the protein was extracted from cell lysates (CL) of K562 and MDA‐MB‐231 cells (MD‐231). a) Intracellular and released L‐plastin, PRDX2 and PRDX4 in K562 and MDA‐MB‐231 cells were assessed by immunoblotting. b) K562 cells were cultured at different cell densities (10 5 –10 6 cells/ml) and the levels of L‐plastin, PRDX2 and PRDX4 in CM were assessed by immunoblotting. Ponceau stain (lower gels) was used as loading control. Shown are representative immunoblots from one out five independent experiments.

Article Snippet: The following Taqman probes were used: L‐plastin, ( Lcp1 , Mm01310735_m1); Peroxiredoxin 2, ( Prdx2 , Mm04208213_m1); Matrix metalloprotease 9, ( Mmp9 , Mm00600163_m1); Trap ( Acp5 , Mm00475698_m1); erythroid transcription factor ( Gata ‐ 1 ; Mm00484679_g1); erythropoietin receptor ( Epor , Mm00438760_m1) and glyceraldehyde 3‐phosphate dehydrogenase ( Gapdh , Mm99999915_g1).

Techniques: Western Blot, Cell Culture, Staining, Control

Journal: Physiological Reports

Article Title: Active hematopoiesis triggers exosomal release of PRDX2 that promotes osteoclast formation

doi: 10.14814/phy2.14745

Figure Lengend Snippet: Acute anemia in mice increase the level of Prdx2 in bone marrow. Acute anemia was induced in female C57BL/6 mice by collecting 10% of total blood volume from saphenous vein, after which groups of animals were sacrificed at days 2, 3, 5 and 7. a‐b) RNA was isolated from bone marrow and gene expression for Prdx2 and Lcp1 was assessed. c‐d) Protein levels of Prdx2 and Lcp1 in bone marrow were assessed by immunoblotting. c) Representative immunoblots for Prdx2 ( top ), Lcp1 ( middle ) and Ponceau S‐stained gel ( bottom ) used for normalization. d) Average protein expression of Prdx2 and L‐plastin. For each immunoblot, protein signal was first normalized to ponceau staining, and then to the maximum signal within the immunoblot. Data are means ±SEM, N = 5–15 mice per condition, * p < 0.05 by one‐way ANOVA with Bonferroni post‐test, red : compared to control

Article Snippet: The following Taqman probes were used: L‐plastin, ( Lcp1 , Mm01310735_m1); Peroxiredoxin 2, ( Prdx2 , Mm04208213_m1); Matrix metalloprotease 9, ( Mmp9 , Mm00600163_m1); Trap ( Acp5 , Mm00475698_m1); erythroid transcription factor ( Gata ‐ 1 ; Mm00484679_g1); erythropoietin receptor ( Epor , Mm00438760_m1) and glyceraldehyde 3‐phosphate dehydrogenase ( Gapdh , Mm99999915_g1).

Techniques: Isolation, Gene Expression, Western Blot, Staining, Expressing, Control

Journal: Physiological Reports

Article Title: Active hematopoiesis triggers exosomal release of PRDX2 that promotes osteoclast formation

doi: 10.14814/phy2.14745

Figure Lengend Snippet: Exosomes from K562 cells induce osteoclast differentiation. a) RAW 264.7 cells were primed with RANKL (50 ng/ml) for 2 days and then cultured for an additional 2 days without RANKL treatment (negative control, NC), with RANKL (50 ng/ml, positive control, PC) or 10% CM from K562 cells +/− exosome inhibitor GW4869 (10 µM). N = 3. b) K562 cells were cultured for 24 h then exosomes were purified, and the distribution of particle sizes was analyzed by Nano sight. c) Representative transmission electron microscopy image of exosomes purified from K562 CM. d‐h) RAW 264.7 cells were primed with RANKL (50 ng/ml) for 2 days and then cultured for an additional 2 days without RANKL treatment (negative control, NC), with RANKL (50 ng/ml, positive control, PC) or with purified exosomes (Exo, 10 µl) from K562 cells. d) Representative images of TRAP‐stained osteoclasts formed in RAW cultures on day 5. Scale bar applies to all images. e) Average osteoclast number; f) average area per osteoclast; g) number of nuclei per osteoclast; h) average area/nucleus. N = 37–50 osteoclasts/condition; i) Immunoblotting for exosomal markers TFR‐2 and TSG101, L‐plastin, PRDX4 and PRDX2 in K562 CM and purified exosomes (Exo). Data are means ±SEM, * p < 0.05, ** p < 0.01 and *** p < 0.001 assessed by Student's t‐test; red : compared to negative control, blue : compared to ‐GW4869 or positive control.

Article Snippet: The following Taqman probes were used: L‐plastin, ( Lcp1 , Mm01310735_m1); Peroxiredoxin 2, ( Prdx2 , Mm04208213_m1); Matrix metalloprotease 9, ( Mmp9 , Mm00600163_m1); Trap ( Acp5 , Mm00475698_m1); erythroid transcription factor ( Gata ‐ 1 ; Mm00484679_g1); erythropoietin receptor ( Epor , Mm00438760_m1) and glyceraldehyde 3‐phosphate dehydrogenase ( Gapdh , Mm99999915_g1).

Techniques: Cell Culture, Negative Control, Positive Control, Purification, Transmission Assay, Electron Microscopy, Staining, Western Blot

Journal: Physiological Reports

Article Title: Active hematopoiesis triggers exosomal release of PRDX2 that promotes osteoclast formation

doi: 10.14814/phy2.14745

Figure Lengend Snippet: Recombinant Prdx2 induces osteoclast differentiation from RANKL‐primed precursors. RAW 264.7 cells were cultured for 4 days without RANKL (white bars), were treated with RANKL (50 ng/ml) for 4 days (black bars) or were RANKL‐primed (treated with RANKL 50 ng/ml for 2 days and cultured for an additional 2 days without RANKL, grey bars). a,b) Prdx2 at concentrations 0.5, 1, 2.5, or 5 µg/ml was added to untreated, RANKL‐treated and RANKL‐primed RAW 264.7. Shown are representative images of TRAP‐positive osteoclasts (a) and average number of osteoclasts formed in different conditions on day 4 (b). Scale bar applies to all images. Data are means ±SD, N = 2–3 replicates. c) Average number of osteoclasts formed when recombinant PRDX2 (2.5 or 5 µg/ml) was added to RANKL‐primed osteoclast precursors. Data are means ±SEM, N = 5–6 independent experiments, * p < 0.05 and ** p < 0.01 compared to RANKL‐primed cultures by Student's t‐test.

Article Snippet: The following Taqman probes were used: L‐plastin, ( Lcp1 , Mm01310735_m1); Peroxiredoxin 2, ( Prdx2 , Mm04208213_m1); Matrix metalloprotease 9, ( Mmp9 , Mm00600163_m1); Trap ( Acp5 , Mm00475698_m1); erythroid transcription factor ( Gata ‐ 1 ; Mm00484679_g1); erythropoietin receptor ( Epor , Mm00438760_m1) and glyceraldehyde 3‐phosphate dehydrogenase ( Gapdh , Mm99999915_g1).

Techniques: Recombinant, Cell Culture

Journal: Cells

Article Title: Conoidin A, a Covalent Inhibitor of Peroxiredoxin 2, Reduces Growth of Glioblastoma Cells by Triggering ROS Production

doi: 10.3390/cells12151934

Figure Lengend Snippet: The level of both PRDX1 and PRDX2 is elevated in GBM tissues and cells. ( A ) The expression of PRDX1 and PRDX2 in NT (n = 20) and GBM (n = 41) tissues. ( B ) Densitometric analysis of PRDX1 and PRDX2 protein bands (relative to β-actin) in NT (n = 20) and GBM (n = 41) tissues. Statistical analysis was performed with Mann–Whitney U test. * p < 0.05. ( C ) Representative blots showing the level of PRDX1 (upper panel) and PRDX2 (lower panel) in NT and GBM tissues. β-actin was used as a loading control. ( D , E ) Representative Western blot results showing the level of PRDX1 ( D ) and PRDX2 ( E ) proteins in NHAs and T98G, U87MG, LN229, LUB17, LUB17N, and LUB20 cells. β-actin was used as a loading control. Bands were quantified by densitometry, determining the quotient of the densitometry signal for PRDX1 or PRDX2 band, and that for β-actin was calculated and then normalized to that of the NHAs. Average fold-change values from three independent protein isolations are shown.

Article Snippet: The quantitative real-time PCR reactions were carried out using the TaqMan Universal PCR Master Mix (Applied Biosystems, Thermo Fisher Scientific) and the primers obtained either from Applied Biosystems (PRDX1, cat. No.: Hs00602020_mH; PRDX2, cat. No.: Hs03044902_g1) or Blirt (Gdansk, PL) ( ACTB coding for β-actin, cat. No.: HK-DD-hu).

Techniques: Expressing, MANN-WHITNEY, Western Blot

Journal: Cell & Bioscience

Article Title: The protective effect of peroxiredoxin II on oxidative stress induced apoptosis in pancreatic β-cells

doi: 10.1186/2045-3701-2-22

Figure Lengend Snippet: PRDX2 is detected in the pancreatic β-cell lines and islet. RT-PCR performed on RNA extracted from INS-1, MIN6, and mouse Islet ( A ). Western blot performed on protein extracted from MIN6, isolated mouse islets, and,mouse pancreas ( B ).

Article Snippet: The cells were then incubated with mouse monoclonal PRDX2 antibody (ABCAM, 1:100) and Alexa 555-conjugated anti-mouse IgG (Jackson Labs, 1:500), and visualized with Nikon Fluorescence microscope (Eclipse TE 200).

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Isolation

Journal: Cell & Bioscience

Article Title: The protective effect of peroxiredoxin II on oxidative stress induced apoptosis in pancreatic β-cells

doi: 10.1186/2045-3701-2-22

Figure Lengend Snippet: PRDX2 expression in β-cells is decreased during oxidative stress induced apoptosis. Western blot performed on proteins extracted from MIN6 cells treated with palmitic acid for indicated concentrations ( A ), or cytokine cocktail for indicated duration ( B ), or STZ for indicated concentrations ( C ). Quantitative analysis of relative PRDX2 expression and relative cleaved caspase 3 expression in MIN6 cells with SEM (A’-C’). n = 3 *p < 0.05.

Article Snippet: The cells were then incubated with mouse monoclonal PRDX2 antibody (ABCAM, 1:100) and Alexa 555-conjugated anti-mouse IgG (Jackson Labs, 1:500), and visualized with Nikon Fluorescence microscope (Eclipse TE 200).

Techniques: Expressing, Western Blot

Journal: Cell & Bioscience

Article Title: The protective effect of peroxiredoxin II on oxidative stress induced apoptosis in pancreatic β-cells

doi: 10.1186/2045-3701-2-22

Figure Lengend Snippet: Elevation of PRDX2 expression protects against oxidative stress induced apoptosis in the β-cells. Western blot of MIN6 cells transfected with PRDX2 or empty vector (EV) and treated with indicated agents ( A ). Quantitative analysis of cleaved caspase 3 expression with SEM, n = 4 *p < 0.05. Immunocytochemistry of MIN6 cells with transfected with PRDX2 (red) or EV (green) and treated with indicated agents. Nuclei of cells were stained with DAPI (blue) ( B ). Quantitative analysis of abnormal nuclear morphology used as an indicator of apoptosis with SEM. n = 3 *p < 0.05. Fluorescence assisted cell sorting (FACS) of MIN6 cells transfected with PRDX2 or EV and treated with indicated agents. Cells were than treated with propidium iodide (PI) only or Annexin V FITC and PI and subjected to FACS. Absorbance in the FL-1 and FL-2 channel was recorded for a minimum of 10,000 events per sample to indicate Annexin V and PI positive cells respectively. ( C ) Quantitative analysis of FL-2 absorbance used as an indicator of cell death with SEM. n = 4, *p < 0.05, **p < 0.01. ( D ) Quantitative analysis of both FL-1 and FL-2 absorbance used as an indicator of apoptosis with SEM. n = 3, *p < 0.05.

Article Snippet: The cells were then incubated with mouse monoclonal PRDX2 antibody (ABCAM, 1:100) and Alexa 555-conjugated anti-mouse IgG (Jackson Labs, 1:500), and visualized with Nikon Fluorescence microscope (Eclipse TE 200).

Techniques: Expressing, Western Blot, Transfection, Plasmid Preparation, Immunocytochemistry, Staining, Fluorescence, FACS

Journal: Cell & Bioscience

Article Title: The protective effect of peroxiredoxin II on oxidative stress induced apoptosis in pancreatic β-cells

doi: 10.1186/2045-3701-2-22

Figure Lengend Snippet: Knockdown of PRDX2 expression exaggerates oxidative stress induced apoptosis in the β-cells. Western blot performed on cell lysate of MIN6 cells with PRDX2 knockdown via siRNA transfection or scrambled control followed with treatment of 0.4 mM palmitic acid or proinflammatory cytokine cocktail for 24 hrs ( A, C ). Quantitative analysis of cleaved caspase 3 expression with SEM, ( B, D ) n = 3 *p < 0.05.

Article Snippet: The cells were then incubated with mouse monoclonal PRDX2 antibody (ABCAM, 1:100) and Alexa 555-conjugated anti-mouse IgG (Jackson Labs, 1:500), and visualized with Nikon Fluorescence microscope (Eclipse TE 200).

Techniques: Expressing, Western Blot, Transfection

Journal: International Journal of Molecular Sciences

Article Title: Upregulation of Phosphorylated HSP27, PRDX2, GRP75, GRP78 and GRP94 in Acquired Middle Ear Cholesteatoma Growth

doi: 10.3390/ijms140714439

Figure Lengend Snippet: Summary of differential proteins identified by LC-MS/MS analysis.

Article Snippet: Rabbit anti-human HSP27, GRP75, GRP78, GRP94 and PRDX2 antibodies were purchased from ProteinTech Group (Chicago, IL, USA).

Techniques: Sequencing, Chemotaxis Assay, Ubiquitin Proteomics, Activation Assay, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Upregulation of Phosphorylated HSP27, PRDX2, GRP75, GRP78 and GRP94 in Acquired Middle Ear Cholesteatoma Growth

doi: 10.3390/ijms140714439

Figure Lengend Snippet: 2D side-by-side comparison of the 2-DE Western blot images of HSP27, PRDX2, GRP78 and GRP75 in cholesteatoma tissues and retroauricular skin run with pI 4–7 and pI 3–10, respectively. Images A , B , C and D are for HSP27. Images E , F , G and H are for PRDX2. Images I , J , K and L are for GRP78. Images M , N , O and P are for GRP75.

Article Snippet: Rabbit anti-human HSP27, GRP75, GRP78, GRP94 and PRDX2 antibodies were purchased from ProteinTech Group (Chicago, IL, USA).

Techniques: Comparison, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Upregulation of Phosphorylated HSP27, PRDX2, GRP75, GRP78 and GRP94 in Acquired Middle Ear Cholesteatoma Growth

doi: 10.3390/ijms140714439

Figure Lengend Snippet: Validation of HSP27, GRP75, GRP78, GRP94 and PRDX2 by Western blotting analysis and RT-PCR. The tissues of cholesteatoma and retroauricular skin were collected from six individual patients. Cholesteatoma is presented by C, and retroauricular skin is presented by S. β-actin was used for normalization.

Article Snippet: Rabbit anti-human HSP27, GRP75, GRP78, GRP94 and PRDX2 antibodies were purchased from ProteinTech Group (Chicago, IL, USA).

Techniques: Biomarker Discovery, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Aging

Article Title: Chetomin rescues pathogenic phenotype of LRRK2 mutation in drosophila.

doi: 10.18632/aging.103843

Figure Lengend Snippet: Figure 1. LRRK2 interacts and phosphorylates PRDX2. (A) LRRK2-GFP and PRDX2-flag are co transfected in HEK293T cells. The lysates were collected and subjected to immunoprecipitation with anti-GFP antibody. The protein was subjected to western blot with anti-flag. Un- transfected cells, LRRK2 GFP alone and PRDX flag alone was transfected into HEK293T cells to serve as negative control. (B) Endogenous colocalization of LRRK2 (green) and PRDX2 (red) protein in SKNSH neuronal cells (C) In vitro kinase assay on SDS-PAGE gel show phosphorylation of PRDX2 by LRRK2 wildtype and G2019S.

Article Snippet: The following antibodies are used: For LRRK2: anti-GFP (Sigma Aldrich G1544), For PRDX2: anti DYKDDDK/Flag (Cell signaling 2368S) and anti PRDX2 (Sigma Aldrich SAB1406520 and EMD Millipore 07-610)

Techniques: Transfection, Immunoprecipitation, Western Blot, Negative Control, In Vitro, Kinase Assay, SDS Page, Phospho-proteomics

Journal: Aging

Article Title: Chetomin rescues pathogenic phenotype of LRRK2 mutation in drosophila.

doi: 10.18632/aging.103843

Figure Lengend Snippet: Figure 2. LRRK2 and PRDX2 transgenic flies. (A) LRRK2 GFP and PRDX2 expression driven by ddc-GAL-4 in transgenic fly head (B) Bar graphs show mean percentage and standard deviation of phosphorylation of PRDX2 in LRRK2 wildtype and G2019S flies (n = 3, cohort of 20).

Article Snippet: The following antibodies are used: For LRRK2: anti-GFP (Sigma Aldrich G1544), For PRDX2: anti DYKDDDK/Flag (Cell signaling 2368S) and anti PRDX2 (Sigma Aldrich SAB1406520 and EMD Millipore 07-610)

Techniques: Transgenic Assay, Expressing, Standard Deviation, Phospho-proteomics

Journal: Aging

Article Title: Chetomin rescues pathogenic phenotype of LRRK2 mutation in drosophila.

doi: 10.18632/aging.103843

Figure Lengend Snippet: Figure 3. PRDX2 is able to rescue G2019S pathogenic phenotype. (A) Representative magnified confocal images of whole mount brains 60 days after eclosion. The different clusters of TH+ neurons are boxed up and labeled. (B) Bar graphs show number of TH-positive DA neuron clusters in flies at 60 days after eclosion (n = 3, cohort of 10). (C) Bar graph shows climbing scores of male flies at 60 days after eclosion. Percentage of flies that reached the top of the column after 1 min was counted (n = 3, cohort of 20). (D) Bar graph shows number of flies that survive after 60 days. Age-matched ddc-GAL4/+ flies were used as controls. Percentage of flies was tabulated. Significance indicated on the graph: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: The following antibodies are used: For LRRK2: anti-GFP (Sigma Aldrich G1544), For PRDX2: anti DYKDDDK/Flag (Cell signaling 2368S) and anti PRDX2 (Sigma Aldrich SAB1406520 and EMD Millipore 07-610)

Techniques: Labeling