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Nacalai blocking solution
Blocking Solution, supplied by Nacalai, 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/product/blocking+solution/blocking+solution/pm42289336-77-8-12
Average 86 stars, based on 1 article reviews
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Article Title: CDKAL1 dysfunction impairs lysine codon translation in podocytes and accelerates chronic kidney disease
Article Snippet: Both cell lines were transduced with lentivirus made by transfecting HEK293FT cells with the sgRNA-containing Lenti-CRISPR v2 Blast, psPAX2 (Addgene), and pMD2.G (Addgene).

Article Title: CDKAL1 dysfunction impairs lysine codon translation in podocytes and accelerates chronic kidney disease.
Article Snippet: Following a 2 h-incubation, the medium in the lower chamber was replaced with RPMI medium supplemented with 40 mg/mL bovine serum albumin (BSA, SigmaAldrich), which had been filtered through a 0.22-μm filter (Millex), and the cells were incubated for an additional 2 h at 37 °C.

Confocal:

Article Title: Spontaneous tauopathy with parkinsonism in an aged cynomolgus macaque
Article Snippet: Antibody binding was visualized using a peroxidase-polymer-based detection system (Histofine Simple Stain MAX-PO kit; Nichirei) with diaminobenzidine as the chromogen. .. For confocal microscopy, sections were similarly processed for antigen retrieval before being incubated in blocking solution (Cat# 06349–64, Nacalai Tesque) for 10 min. .. This was followed by overnight incubation at 4 °C with primary antibodies, and then a 1-h incubation at room temperature with Alexa Fluor Plus 488, 594, or 647-conjugated secondary antibodies (Thermo Fisher Scientific).

Microscopy:

Article Title: Spontaneous tauopathy with parkinsonism in an aged cynomolgus macaque
Article Snippet: Antibody binding was visualized using a peroxidase-polymer-based detection system (Histofine Simple Stain MAX-PO kit; Nichirei) with diaminobenzidine as the chromogen. .. For confocal microscopy, sections were similarly processed for antigen retrieval before being incubated in blocking solution (Cat# 06349–64, Nacalai Tesque) for 10 min. .. This was followed by overnight incubation at 4 °C with primary antibodies, and then a 1-h incubation at room temperature with Alexa Fluor Plus 488, 594, or 647-conjugated secondary antibodies (Thermo Fisher Scientific).

Incubation:

Article Title: Spontaneous tauopathy with parkinsonism in an aged cynomolgus macaque
Article Snippet: Antibody binding was visualized using a peroxidase-polymer-based detection system (Histofine Simple Stain MAX-PO kit; Nichirei) with diaminobenzidine as the chromogen. .. For confocal microscopy, sections were similarly processed for antigen retrieval before being incubated in blocking solution (Cat# 06349–64, Nacalai Tesque) for 10 min. .. This was followed by overnight incubation at 4 °C with primary antibodies, and then a 1-h incubation at room temperature with Alexa Fluor Plus 488, 594, or 647-conjugated secondary antibodies (Thermo Fisher Scientific).

Article Title: Optimum Serum Concentration Enhances Migration of MDA-MB-231 Triple-Negative Breast Cancer Cells and Promotes Intracellular Delivery of Proapoptotic Domain via Cell-Penetrating Peptides
Article Snippet: .. Samples were blocked with blocking solution (Nakalai Tesque) and incubated for 60 min at 25 °C. .. Nuclear and actin staining was performed using 300 nM DAPI and Alexa Fluor 488 Phalloidin (1:200, Invitrogen) in PBS for 20 min at 25 °C, followed by three 20 min rinses with PBS.

Article Title: Role of dynamic changes in human sperm heat shock protein A2 in male infertility.
Article Snippet: .. After washing, the slides were 145 blocked with blocking solution (Blocking One-P; Nacalai Tesque, Inc., Kyoto, Japan) and incubated for 146 1 h at room temperature. ..

Blocking Assay:

Article Title: Spontaneous tauopathy with parkinsonism in an aged cynomolgus macaque
Article Snippet: Antibody binding was visualized using a peroxidase-polymer-based detection system (Histofine Simple Stain MAX-PO kit; Nichirei) with diaminobenzidine as the chromogen. .. For confocal microscopy, sections were similarly processed for antigen retrieval before being incubated in blocking solution (Cat# 06349–64, Nacalai Tesque) for 10 min. .. This was followed by overnight incubation at 4 °C with primary antibodies, and then a 1-h incubation at room temperature with Alexa Fluor Plus 488, 594, or 647-conjugated secondary antibodies (Thermo Fisher Scientific).

Article Title: Optimum Serum Concentration Enhances Migration of MDA-MB-231 Triple-Negative Breast Cancer Cells and Promotes Intracellular Delivery of Proapoptotic Domain via Cell-Penetrating Peptides
Article Snippet: .. Samples were blocked with blocking solution (Nakalai Tesque) and incubated for 60 min at 25 °C. .. Nuclear and actin staining was performed using 300 nM DAPI and Alexa Fluor 488 Phalloidin (1:200, Invitrogen) in PBS for 20 min at 25 °C, followed by three 20 min rinses with PBS.

Article Title: High CD36 Expression Predicts Aggressive Invasion and Recurrence in Oral Squamous Cell Carcinoma
Article Snippet: .. To prevent nonspecific binding of the antibodies, the sections were immersed in a blocking solution (Nacalai Tesque, Kyoto, Japan) for 15 min. .. Subsequently, the sections were incubated with the primary antibody (anti-CD36 rabbit monoclonal antibody, 1:100; clone EPR6573; Abcam, Cambridge, UK) overnight at 4 °C.

Article Title: Selenoprotein P deletion ameliorates metabolic stress-associated anxiety-like behavior in male mice.
Article Snippet: .. RNAscope ISH) After RNAscope ISH, sections were permeabilized with 0.3% Triton X-100 in PBS and treated with a blocking solution (Blocking one histo, Nacalai Tesque, #L2E9350) containing 0.3% Triton X-100 in PBS. .. The sections were incubated overnight at 4 °C with anti-GS (Abcam, Cat# ab228590, rabbit, 1:500, RRID: AB_3665798), anti-Iba1 (Wako, Cat# 019-19741, RRID: AB_839504, rabbit, 1:1000), or NeuN (Cell Signaling Technology Cat# 12943, RRID: AB_2630395) primary antibodies and washed in PBS.

Article Title: Role of dynamic changes in human sperm heat shock protein A2 in male infertility.
Article Snippet: .. After washing, the slides were 145 blocked with blocking solution (Blocking One-P; Nacalai Tesque, Inc., Kyoto, Japan) and incubated for 146 1 h at room temperature. ..

Article Title: CDKAL1 dysfunction impairs lysine codon translation in podocytes and accelerates chronic kidney disease
Article Snippet: O.C.T.Compund , Tissue–Tek , 4583. .. Blocking solution , Nacalai Tasque , 06349-64. .. Dual-Luciferase Reporter Assay System , Promega , E1910.

Binding Assay:

Article Title: High CD36 Expression Predicts Aggressive Invasion and Recurrence in Oral Squamous Cell Carcinoma
Article Snippet: .. To prevent nonspecific binding of the antibodies, the sections were immersed in a blocking solution (Nacalai Tesque, Kyoto, Japan) for 15 min. .. Subsequently, the sections were incubated with the primary antibody (anti-CD36 rabbit monoclonal antibody, 1:100; clone EPR6573; Abcam, Cambridge, UK) overnight at 4 °C.

RNAscope:

Article Title: Selenoprotein P deletion ameliorates metabolic stress-associated anxiety-like behavior in male mice.
Article Snippet: .. RNAscope ISH) After RNAscope ISH, sections were permeabilized with 0.3% Triton X-100 in PBS and treated with a blocking solution (Blocking one histo, Nacalai Tesque, #L2E9350) containing 0.3% Triton X-100 in PBS. .. The sections were incubated overnight at 4 °C with anti-GS (Abcam, Cat# ab228590, rabbit, 1:500, RRID: AB_3665798), anti-Iba1 (Wako, Cat# 019-19741, RRID: AB_839504, rabbit, 1:1000), or NeuN (Cell Signaling Technology Cat# 12943, RRID: AB_2630395) primary antibodies and washed in PBS.

In Situ Hybridization:

Article Title: Selenoprotein P deletion ameliorates metabolic stress-associated anxiety-like behavior in male mice.
Article Snippet: .. RNAscope ISH) After RNAscope ISH, sections were permeabilized with 0.3% Triton X-100 in PBS and treated with a blocking solution (Blocking one histo, Nacalai Tesque, #L2E9350) containing 0.3% Triton X-100 in PBS. .. The sections were incubated overnight at 4 °C with anti-GS (Abcam, Cat# ab228590, rabbit, 1:500, RRID: AB_3665798), anti-Iba1 (Wako, Cat# 019-19741, RRID: AB_839504, rabbit, 1:1000), or NeuN (Cell Signaling Technology Cat# 12943, RRID: AB_2630395) primary antibodies and washed in PBS.



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Expression and localization of <t>CA2</t> in the human placentas and Serum. (A) Heatmap of carbonic anhydrase family expression in placental transcriptomes from normal and preeclamptic pregnancies. (B) Box plot analysis of carbonic anhydrase family expression in placental transcriptomes from normal and preeclamptic pregnancies. (C) MRNA expression level of CA2 in PE and normal placentas detected by RT‐qPCR in the discovery cohort. (D) Representative western blot bands for CA2. (E) Protein expression level of CA2 in PE and normal placentas detected by Western Blot in the discovery cohort. (F) Serum expression level of CA2 in PE and normal women detected by ELISA in the serum validation cohort. (G) Immunohistochemistry method to detect the expression localization of CA2 in placentas from normal and preeclamptic pregnancies. Scale bars: 3 mm (overview), 200 μm (magnified field). All quantitative data are expressed as means ± SD; ns: p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.
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Expression and localization of <t>CA2</t> in the human placentas and Serum. (A) Heatmap of carbonic anhydrase family expression in placental transcriptomes from normal and preeclamptic pregnancies. (B) Box plot analysis of carbonic anhydrase family expression in placental transcriptomes from normal and preeclamptic pregnancies. (C) MRNA expression level of CA2 in PE and normal placentas detected by RT‐qPCR in the discovery cohort. (D) Representative western blot bands for CA2. (E) Protein expression level of CA2 in PE and normal placentas detected by Western Blot in the discovery cohort. (F) Serum expression level of CA2 in PE and normal women detected by ELISA in the serum validation cohort. (G) Immunohistochemistry method to detect the expression localization of CA2 in placentas from normal and preeclamptic pregnancies. Scale bars: 3 mm (overview), 200 μm (magnified field). All quantitative data are expressed as means ± SD; ns: p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.
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Expression and localization of <t>CA2</t> in the human placentas and Serum. (A) Heatmap of carbonic anhydrase family expression in placental transcriptomes from normal and preeclamptic pregnancies. (B) Box plot analysis of carbonic anhydrase family expression in placental transcriptomes from normal and preeclamptic pregnancies. (C) MRNA expression level of CA2 in PE and normal placentas detected by RT‐qPCR in the discovery cohort. (D) Representative western blot bands for CA2. (E) Protein expression level of CA2 in PE and normal placentas detected by Western Blot in the discovery cohort. (F) Serum expression level of CA2 in PE and normal women detected by ELISA in the serum validation cohort. (G) Immunohistochemistry method to detect the expression localization of CA2 in placentas from normal and preeclamptic pregnancies. Scale bars: 3 mm (overview), 200 μm (magnified field). All quantitative data are expressed as means ± SD; ns: p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.
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Expression and localization of CA2 in the human placentas and Serum. (A) Heatmap of carbonic anhydrase family expression in placental transcriptomes from normal and preeclamptic pregnancies. (B) Box plot analysis of carbonic anhydrase family expression in placental transcriptomes from normal and preeclamptic pregnancies. (C) MRNA expression level of CA2 in PE and normal placentas detected by RT‐qPCR in the discovery cohort. (D) Representative western blot bands for CA2. (E) Protein expression level of CA2 in PE and normal placentas detected by Western Blot in the discovery cohort. (F) Serum expression level of CA2 in PE and normal women detected by ELISA in the serum validation cohort. (G) Immunohistochemistry method to detect the expression localization of CA2 in placentas from normal and preeclamptic pregnancies. Scale bars: 3 mm (overview), 200 μm (magnified field). All quantitative data are expressed as means ± SD; ns: p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.

Journal: The FASEB Journal

Article Title: Hypoxia‐Driven CA2 Promotes Trophoblast Dysfunction in Preeclampsia

doi: 10.1096/fj.202602024R

Figure Lengend Snippet: Expression and localization of CA2 in the human placentas and Serum. (A) Heatmap of carbonic anhydrase family expression in placental transcriptomes from normal and preeclamptic pregnancies. (B) Box plot analysis of carbonic anhydrase family expression in placental transcriptomes from normal and preeclamptic pregnancies. (C) MRNA expression level of CA2 in PE and normal placentas detected by RT‐qPCR in the discovery cohort. (D) Representative western blot bands for CA2. (E) Protein expression level of CA2 in PE and normal placentas detected by Western Blot in the discovery cohort. (F) Serum expression level of CA2 in PE and normal women detected by ELISA in the serum validation cohort. (G) Immunohistochemistry method to detect the expression localization of CA2 in placentas from normal and preeclamptic pregnancies. Scale bars: 3 mm (overview), 200 μm (magnified field). All quantitative data are expressed as means ± SD; ns: p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: After PBS washing, nonspecific binding sites were blocked with 5% bovine serum albumin (BSA) at room temperature for 1 h. Primary antibody incubation: Excess blocking solution was discarded, and sections were incubated with primary antibody targeting CA2 (Cat. 16961‐AP, Proteintech, Wuhan, China) overnight at 4°C in a humidified dark box.

Techniques: Expressing, Analysis, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Biomarker Discovery, Immunohistochemistry, Reverse Transcription Polymerase Chain Reaction

Correlation analysis between CA2 expression and clinical indicators. (A) Correlation of CA2 levels with SBP. (B) Correlation of CA2 levels with DBP. (C) Correlation of CA2 levels with Fetal Weight. (D) Correlation of CA2 levels with APGAR‐1 min. (E) Correlation of CA2 levels with APGAR‐5 min. (F) Correlation of CA2 levels with Proteinuria. (G) Correlation of CA2 levels with maternal blood creatinine levels before delivery. (H) Correlation of CA2 levels with blood urea levels. (I) Correlation of CA2 levels with ALT. (J) Correlation of CA2 levels with AST. (K) Correlation of CA2 levels with PT levels. (L) Correlation of CA2 levels with APTT. (M) Correlation of CA2 levels with platelet levels. Correlation analyses were performed using Pearson's correlation coefficient. ns: p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.

Journal: The FASEB Journal

Article Title: Hypoxia‐Driven CA2 Promotes Trophoblast Dysfunction in Preeclampsia

doi: 10.1096/fj.202602024R

Figure Lengend Snippet: Correlation analysis between CA2 expression and clinical indicators. (A) Correlation of CA2 levels with SBP. (B) Correlation of CA2 levels with DBP. (C) Correlation of CA2 levels with Fetal Weight. (D) Correlation of CA2 levels with APGAR‐1 min. (E) Correlation of CA2 levels with APGAR‐5 min. (F) Correlation of CA2 levels with Proteinuria. (G) Correlation of CA2 levels with maternal blood creatinine levels before delivery. (H) Correlation of CA2 levels with blood urea levels. (I) Correlation of CA2 levels with ALT. (J) Correlation of CA2 levels with AST. (K) Correlation of CA2 levels with PT levels. (L) Correlation of CA2 levels with APTT. (M) Correlation of CA2 levels with platelet levels. Correlation analyses were performed using Pearson's correlation coefficient. ns: p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: After PBS washing, nonspecific binding sites were blocked with 5% bovine serum albumin (BSA) at room temperature for 1 h. Primary antibody incubation: Excess blocking solution was discarded, and sections were incubated with primary antibody targeting CA2 (Cat. 16961‐AP, Proteintech, Wuhan, China) overnight at 4°C in a humidified dark box.

Techniques: Analysis, Expressing

Preeclampsia modeling and placental CA2 expression in L‐NAME mouse model. (A) Experimental workflow of the L‐NAME‐induced preeclampsia model. (B) Changes in systolic blood pressure in L‐NAME and CONTROL groups. (C, E) Body weights of fetal mice in L‐NAME and CONTROL groups. (D) Placental efficiency of fetal mice in L‐NAME and CONTROL groups. (H) SFLT‐1 levels of mice in L‐NAME and CONTROL groups. (F) The ratio of urinary albumin and creatine of mice in L‐NAME and CONTROL groups. (G, I) Representative renal and placental stained with hematoxylin & eosin (H&E) in L‐NAME and CONTROL groups. Scale bars: 250 μm. (J) Ratio of palcental labyrinth and junction zone of mice in L‐NAME and CONTROL groups. (K) MRNA expression level of placental CA2 in L‐NAME and CONTROL groups detected by RT‐qPCR. (L) Representative western blot bands for CA2. (M) Protein expression level of placental CA2 in L‐NAME and CONTROL groups detected by Western Blot. All quantitative data are expressed as means ± SD; ns: p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.

Journal: The FASEB Journal

Article Title: Hypoxia‐Driven CA2 Promotes Trophoblast Dysfunction in Preeclampsia

doi: 10.1096/fj.202602024R

Figure Lengend Snippet: Preeclampsia modeling and placental CA2 expression in L‐NAME mouse model. (A) Experimental workflow of the L‐NAME‐induced preeclampsia model. (B) Changes in systolic blood pressure in L‐NAME and CONTROL groups. (C, E) Body weights of fetal mice in L‐NAME and CONTROL groups. (D) Placental efficiency of fetal mice in L‐NAME and CONTROL groups. (H) SFLT‐1 levels of mice in L‐NAME and CONTROL groups. (F) The ratio of urinary albumin and creatine of mice in L‐NAME and CONTROL groups. (G, I) Representative renal and placental stained with hematoxylin & eosin (H&E) in L‐NAME and CONTROL groups. Scale bars: 250 μm. (J) Ratio of palcental labyrinth and junction zone of mice in L‐NAME and CONTROL groups. (K) MRNA expression level of placental CA2 in L‐NAME and CONTROL groups detected by RT‐qPCR. (L) Representative western blot bands for CA2. (M) Protein expression level of placental CA2 in L‐NAME and CONTROL groups detected by Western Blot. All quantitative data are expressed as means ± SD; ns: p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: After PBS washing, nonspecific binding sites were blocked with 5% bovine serum albumin (BSA) at room temperature for 1 h. Primary antibody incubation: Excess blocking solution was discarded, and sections were incubated with primary antibody targeting CA2 (Cat. 16961‐AP, Proteintech, Wuhan, China) overnight at 4°C in a humidified dark box.

Techniques: Expressing, Control, Staining, Quantitative RT-PCR, Western Blot, Reverse Transcription Polymerase Chain Reaction

Effects of CA2 on trophoblast cell function. (A) RT‐qPCR analysis to detect the expression of mRNA level after overexpression of CA2 in HTR‐8/SVneo cells. (B) Representative western blot bands for CA2. (C) Western Blot analysis to detect the expression of protein level after overexpression of CA2 in HTR‐8/SVneo cells. (D) Effect of CA2 overexpression on the proliferative capacity of HTR‐8/SVneo cells. (E) Impact of CA2 overexpression on the migration and invasion abilities of HTR‐8/SVneo cells. (F) Statistical analysis of the Transwell migration and invasion assays. (G) Effect of CA2 overexpression on the apoptotic properties of HTR‐8/SVneo cells. (H) Statistical analysis of the apoptosis assay. pEX‐3‐control: Empty vector control group; pEX‐3‐CA2: CA2 overexpression group. All quantitative data are expressed as means ± SD; ns: p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.

Journal: The FASEB Journal

Article Title: Hypoxia‐Driven CA2 Promotes Trophoblast Dysfunction in Preeclampsia

doi: 10.1096/fj.202602024R

Figure Lengend Snippet: Effects of CA2 on trophoblast cell function. (A) RT‐qPCR analysis to detect the expression of mRNA level after overexpression of CA2 in HTR‐8/SVneo cells. (B) Representative western blot bands for CA2. (C) Western Blot analysis to detect the expression of protein level after overexpression of CA2 in HTR‐8/SVneo cells. (D) Effect of CA2 overexpression on the proliferative capacity of HTR‐8/SVneo cells. (E) Impact of CA2 overexpression on the migration and invasion abilities of HTR‐8/SVneo cells. (F) Statistical analysis of the Transwell migration and invasion assays. (G) Effect of CA2 overexpression on the apoptotic properties of HTR‐8/SVneo cells. (H) Statistical analysis of the apoptosis assay. pEX‐3‐control: Empty vector control group; pEX‐3‐CA2: CA2 overexpression group. All quantitative data are expressed as means ± SD; ns: p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: After PBS washing, nonspecific binding sites were blocked with 5% bovine serum albumin (BSA) at room temperature for 1 h. Primary antibody incubation: Excess blocking solution was discarded, and sections were incubated with primary antibody targeting CA2 (Cat. 16961‐AP, Proteintech, Wuhan, China) overnight at 4°C in a humidified dark box.

Techniques: Cell Function Assay, Quantitative RT-PCR, Analysis, Expressing, Over Expression, Western Blot, Migration, Apoptosis Assay, Control, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction

VEGFA overexpression rescues the impaired proliferation, migration and invasion of HTR‐8/SVneo cells induced by CA2 overexpression. (A) Heatmap showing hierarchical clustering of differentially expressed genes between the control and CA2 overexpression groups. (B) Volcano plot of differentially expressed genes. (C) Western Blot analysis of CA2 and VEGFA to detect the protein expression level on the of HTR‐8/SVneo cells in each expremental group. (D) Proliferative capacity analysis of HTR‐8/SVneo cells in the control group、CA2 overexpression group and CA2 and VEGFA co‐overexpression group. (E) Migration and invasion abilities of HTR‐228/SVneo cells in the control group, CA2 overexpression group and CA2 and VEGFA co‐overexpression group. (F) Statistical analysis of the Transwell migration and invasion abilities in the control group, CA2 overexpression group and CA2 and VEGFA co‐overexpression group. pEX‐3‐control 1/2/3: Empty vector control group; pEX‐3‐CA2 1/2/3: CA2 overexpression group; pEX‐3‐control: Empty vector control group; pEX‐3‐ CA2: CA2 overexpression group; pEX‐3‐ CA2+ pEX‐3‐ VEGFA: CA2 and VEGFA co‐overexpression group. All quantitative data are expressed as means ± SD; ns: p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.

Journal: The FASEB Journal

Article Title: Hypoxia‐Driven CA2 Promotes Trophoblast Dysfunction in Preeclampsia

doi: 10.1096/fj.202602024R

Figure Lengend Snippet: VEGFA overexpression rescues the impaired proliferation, migration and invasion of HTR‐8/SVneo cells induced by CA2 overexpression. (A) Heatmap showing hierarchical clustering of differentially expressed genes between the control and CA2 overexpression groups. (B) Volcano plot of differentially expressed genes. (C) Western Blot analysis of CA2 and VEGFA to detect the protein expression level on the of HTR‐8/SVneo cells in each expremental group. (D) Proliferative capacity analysis of HTR‐8/SVneo cells in the control group、CA2 overexpression group and CA2 and VEGFA co‐overexpression group. (E) Migration and invasion abilities of HTR‐228/SVneo cells in the control group, CA2 overexpression group and CA2 and VEGFA co‐overexpression group. (F) Statistical analysis of the Transwell migration and invasion abilities in the control group, CA2 overexpression group and CA2 and VEGFA co‐overexpression group. pEX‐3‐control 1/2/3: Empty vector control group; pEX‐3‐CA2 1/2/3: CA2 overexpression group; pEX‐3‐control: Empty vector control group; pEX‐3‐ CA2: CA2 overexpression group; pEX‐3‐ CA2+ pEX‐3‐ VEGFA: CA2 and VEGFA co‐overexpression group. All quantitative data are expressed as means ± SD; ns: p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: After PBS washing, nonspecific binding sites were blocked with 5% bovine serum albumin (BSA) at room temperature for 1 h. Primary antibody incubation: Excess blocking solution was discarded, and sections were incubated with primary antibody targeting CA2 (Cat. 16961‐AP, Proteintech, Wuhan, China) overnight at 4°C in a humidified dark box.

Techniques: Over Expression, Migration, Control, Western Blot, Analysis, Expressing, Plasmid Preparation

HIF‐1α activates the transcription of CA2. (A, B) RT‐qPCR analysis of CA2 and HIF‐1α to detect the expression of mRNA level under hypoxic or normoxia exposure in HTR‐8/Svneo cells. (C) Representative western blot bands. (D, E) Western Blot analysis of CA2 and HIF‐1α to detect the expression of protein level under hypoxic or normoxia exposure in HTR‐8/Svneo cells. (F, G) RT‐qPCR analysis of CA2 and HIF‐1α to detect the expression of mRNA level after overexpression HIF‐1α in HTR‐8/Svneo cells. (H) Representative western blot bands. (I, J) Western Blot analysis of CA2 and HIF‐1α to detect the expression of protein level after overexpression HIF‐1α in HTR‐8/Svneo cells. (K) Luciferase activity of HTR‐8/SVneo cells with or without overexpression HIF‐1α. (L) Luciferase activity of HTR‐8/SVneo cells with or without exposing hypoxic environment. pcDNA3.1‐control: Empty vector control group; pcDNA3.1‐HIF‐1α: HIF‐1α overexpression group; CA2 promoter + Vecter: Plasmid with CA2 promoter + empty vector control group; CA2 promoter + HIF‐1α OE: Plasmid with CA2 promoter + HIF‐1α overexpression group; CA2 promoter + Vecter + hypoxia 12 h: Plasmid with CA2 promoter + empty vector control group + exposing hypoxic environment 12 h; CA2 promoter + Vecter + hypoxia 24 h: Plasmid with CA2 promoter + empty vector control group + exposing hypoxic environment 24 h; CA2 promoter + Vecter + hypoxia 36 h: Plasmid with CA2 promoter + empty vector control group + exposing hypoxic environment 36 h. All quantitative data are expressed as means ± SD; ns: p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.

Journal: The FASEB Journal

Article Title: Hypoxia‐Driven CA2 Promotes Trophoblast Dysfunction in Preeclampsia

doi: 10.1096/fj.202602024R

Figure Lengend Snippet: HIF‐1α activates the transcription of CA2. (A, B) RT‐qPCR analysis of CA2 and HIF‐1α to detect the expression of mRNA level under hypoxic or normoxia exposure in HTR‐8/Svneo cells. (C) Representative western blot bands. (D, E) Western Blot analysis of CA2 and HIF‐1α to detect the expression of protein level under hypoxic or normoxia exposure in HTR‐8/Svneo cells. (F, G) RT‐qPCR analysis of CA2 and HIF‐1α to detect the expression of mRNA level after overexpression HIF‐1α in HTR‐8/Svneo cells. (H) Representative western blot bands. (I, J) Western Blot analysis of CA2 and HIF‐1α to detect the expression of protein level after overexpression HIF‐1α in HTR‐8/Svneo cells. (K) Luciferase activity of HTR‐8/SVneo cells with or without overexpression HIF‐1α. (L) Luciferase activity of HTR‐8/SVneo cells with or without exposing hypoxic environment. pcDNA3.1‐control: Empty vector control group; pcDNA3.1‐HIF‐1α: HIF‐1α overexpression group; CA2 promoter + Vecter: Plasmid with CA2 promoter + empty vector control group; CA2 promoter + HIF‐1α OE: Plasmid with CA2 promoter + HIF‐1α overexpression group; CA2 promoter + Vecter + hypoxia 12 h: Plasmid with CA2 promoter + empty vector control group + exposing hypoxic environment 12 h; CA2 promoter + Vecter + hypoxia 24 h: Plasmid with CA2 promoter + empty vector control group + exposing hypoxic environment 24 h; CA2 promoter + Vecter + hypoxia 36 h: Plasmid with CA2 promoter + empty vector control group + exposing hypoxic environment 36 h. All quantitative data are expressed as means ± SD; ns: p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: After PBS washing, nonspecific binding sites were blocked with 5% bovine serum albumin (BSA) at room temperature for 1 h. Primary antibody incubation: Excess blocking solution was discarded, and sections were incubated with primary antibody targeting CA2 (Cat. 16961‐AP, Proteintech, Wuhan, China) overnight at 4°C in a humidified dark box.

Techniques: Quantitative RT-PCR, Analysis, Expressing, Western Blot, Over Expression, Luciferase, Activity Assay, Control, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction

Inhibition of CA2 partially improves outcomes in a L‐NAME‐induced PE mouse model. (A) Experimental workflow for LNP‐NC and LNP‐siCA2 treatment in L‐NAME PE mice. (B) MRNA expression level of placental CA2 in LNP‐NC and LNP‐siCA2 groups detected by RT‐qPCR. (C) Representative immunofluorescence staining of CA2 in LNP‐NC and LNP‐siCA2 groups. Scale bars: 500 μm. (D) Quantitative analysis of mean fluorescence intensity of CA2 immunofluorescence signals n LNP‐NC and LNP‐siCA2 groups. (E) Changes in systolic blood pressure in LNP‐NC and LNP‐siCA2 groups. (F) The ratio of urinary albumin and creatine of mice in LNP‐NC group and LNP‐siCA2 group. (G) Representative renal stained with hematoxylin & eosin (H&E) in LNP‐NC and LNP‐siCA2 groups. (H) SFLT‐1 levels of mice in LNP‐NC and LNP‐siCA2 groups. (I, K) Body weights of fetal mice in LNP‐NC and LNP‐siCA2 groups. (J) Placental efficiency of fetal mice in LNP‐NC group and LNP‐siCA2 group. (L) Representative placental stained with hematoxylin & eosin (H&E) in LNP‐NC and LNP‐siCA2 groups. Scale bars: 250 μm. (M) Ratio of palcental labyrinth and junction zone of mice in LNP‐NC and LNP‐siCA2 groups. All quantitative data are expressed as means ± SD; ns: p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.

Journal: The FASEB Journal

Article Title: Hypoxia‐Driven CA2 Promotes Trophoblast Dysfunction in Preeclampsia

doi: 10.1096/fj.202602024R

Figure Lengend Snippet: Inhibition of CA2 partially improves outcomes in a L‐NAME‐induced PE mouse model. (A) Experimental workflow for LNP‐NC and LNP‐siCA2 treatment in L‐NAME PE mice. (B) MRNA expression level of placental CA2 in LNP‐NC and LNP‐siCA2 groups detected by RT‐qPCR. (C) Representative immunofluorescence staining of CA2 in LNP‐NC and LNP‐siCA2 groups. Scale bars: 500 μm. (D) Quantitative analysis of mean fluorescence intensity of CA2 immunofluorescence signals n LNP‐NC and LNP‐siCA2 groups. (E) Changes in systolic blood pressure in LNP‐NC and LNP‐siCA2 groups. (F) The ratio of urinary albumin and creatine of mice in LNP‐NC group and LNP‐siCA2 group. (G) Representative renal stained with hematoxylin & eosin (H&E) in LNP‐NC and LNP‐siCA2 groups. (H) SFLT‐1 levels of mice in LNP‐NC and LNP‐siCA2 groups. (I, K) Body weights of fetal mice in LNP‐NC and LNP‐siCA2 groups. (J) Placental efficiency of fetal mice in LNP‐NC group and LNP‐siCA2 group. (L) Representative placental stained with hematoxylin & eosin (H&E) in LNP‐NC and LNP‐siCA2 groups. Scale bars: 250 μm. (M) Ratio of palcental labyrinth and junction zone of mice in LNP‐NC and LNP‐siCA2 groups. All quantitative data are expressed as means ± SD; ns: p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: After PBS washing, nonspecific binding sites were blocked with 5% bovine serum albumin (BSA) at room temperature for 1 h. Primary antibody incubation: Excess blocking solution was discarded, and sections were incubated with primary antibody targeting CA2 (Cat. 16961‐AP, Proteintech, Wuhan, China) overnight at 4°C in a humidified dark box.

Techniques: Inhibition, Expressing, Quantitative RT-PCR, Immunofluorescence, Staining, Analysis, Fluorescence, Reverse Transcription Polymerase Chain Reaction