human aldoa antibody  (Proteintech)

Journal: OncoTargets and Therapy

Article Title: The Expression and Clinical Significance of ALDOA in Breast Cancer

doi: 10.2147/OTT.S518473

Figure Lengend Snippet: ALDOA expression in human BC tissues. ( A ) ALDOA expression in tumor and normal tissues in TIMER database. ( B – D ) ALDOA expression in BC tumor and normal tissues in bc‐GenExMiner v5.1 ( B ), UALCAN ( C ) and GEPIA ( D ) databases. ( E ) Representative IHC staining of ALDOA in human BC tissues and normal tissues. ( F ) Analysis of ALDOA IHC scores in human BC tissues and normal tissues. *** P < 0.001.

Article Snippet: Following defined procedures, the paraffin-embedded tissues were sectioned to a thickness of 5 μm, incubated at 4°C overnight with a monoclonal human ALDOA antibody (dilution 1:100; #11305-1-AP, Proteintech), stained using a staining kit (Zhongshan Biotechnology, Bei-jing, China), followed by visualization.

Techniques: Expressing, Immunohistochemistry

Journal: Cell death discovery

Article Title: ALDOA contributes to colorectal tumorigenesis and metastasis by targeting YAP.

doi: 10.1038/s41420-024-02249-z

Figure Lengend Snippet: Fig. 5 ALDOA modulates YAP activity. A Western blotting analysis of p-YAP and YAP expression in HT-29 and DLD-1 cells (NC vs. KD). B Western blotting analysis of p-YAP and YAP expression in SW480 and SW620 cells (VEC vs. OE). C Subcellular fractionation analysis of YAP expression in HT-29 cells (NC vs. KD). GAPDH served as control for the cytoplasmic (C) fraction, and Lamin B served as control for the nuclear (N) fractions. D IF staining of YAP (red) in HT-29 cells (NC vs. KD). Nuclei were stained with DAPI (blue). E Typical images of IHC staining of ALDOA and AREG in the xenograft tumors derived from HT-29 cells (NC vs. KD). F Western blotting analysis of the indicated proteins in the xenograft tumors derived from HT-29 cells (NC vs. KD). (G qRT-PCR analysis of CTGF and AREG mRNA levels in HT-29 cells (NC vs. KD). H qRT- PCR analysis of CTGF and AREG mRNA levels in SW480 cells (VEC vs. OE). *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: In this study, the primary antibodies detecting human ALDOA, AREG and CTGF (dilution 1:100; Proteintech) were used.

Techniques: Activity Assay, Western Blot, Expressing, Fractionation, Control, Staining, Immunohistochemistry, Derivative Assay, Quantitative RT-PCR

Journal: Cell death discovery

Article Title: ALDOA contributes to colorectal tumorigenesis and metastasis by targeting YAP.

doi: 10.1038/s41420-024-02249-z

Figure Lengend Snippet: Fig. 7 ALDOA regulates YAP activity through AMPK pathway. A Western blot analysis of p-AMPK and AMPK expression in HT-29 and DLD-1 cells (NC vs. KD). B Western blot analysis of p-AMPK and AMPK expression in SW480 and SW620 cells (VEC vs. OE). C Western blotting analysis of the indicated proteins in HT-29 cells (NC vs. KD) treated with siRNA against AMPK (siAMPK) and siControl. D qRT-PCR analysis of CTGF mRNA expression in SW480 cells (VEC vs. OE) treated with siAMPK and siControl. E qRT-PCR analysis of AREG mRNA expression in SW480 cells (VEC vs. OE) treated with siAMPK and siControl. ns, nonsignificant, **P < 0.01, ***P < 0.001.

Article Snippet: In this study, the primary antibodies detecting human ALDOA, AREG and CTGF (dilution 1:100; Proteintech) were used.

Techniques: Activity Assay, Western Blot, Expressing, Quantitative RT-PCR

Journal: Cell death discovery

Article Title: ALDOA contributes to colorectal tumorigenesis and metastasis by targeting YAP.

doi: 10.1038/s41420-024-02249-z

Figure Lengend Snippet: Fig. 8 The expression levels of ALDOA, CTGF and AREG in clinical CRC tissues. A IHC staining of ALDOA, CTGF and AREG in CRC tumor tissues. Spearman correlation analysis between ALDOA and CTGF (B), and ALDOA and AREG (C) in CRC tumor tissues. D IHC staining of ALDOA, CTGF and AREG in normal tissues. Spearman correlation analysis between ALDOA and CTGF (E), and ALDOA and AREG (F) in normal tissues. G Correlation analysis of ALDOA and CTGF gene expression based on the published microarray dataset (GSE37182). H Correlation analysis of ALDOA and AREG gene expression based on the published microarray dataset (GSE37182). I Correlation analysis of ALDOA and CTGF gene expression based on the published microarray dataset (GSE73360). J Correlation analysis of ALDOA and AREG gene expression based on the published microarray dataset (GSE73360).

Article Snippet: In this study, the primary antibodies detecting human ALDOA, AREG and CTGF (dilution 1:100; Proteintech) were used.

Techniques: Expressing, Immunohistochemistry, Gene Expression, Microarray

Journal: Cell death discovery

Article Title: ALDOA contributes to colorectal tumorigenesis and metastasis by targeting YAP.

doi: 10.1038/s41420-024-02249-z

Figure Lengend Snippet: Fig. 10 Schematic diagram of the molecular mechanism of ALDOA regulation of AMPK/YAP pathway. High ALDOA expression leads to AMPK inhibition and YAP unphosphorylation. Unphophorylated YAP translocates into the nucleus and functions as a transcriptional co- activator by binding to the TEAD family of transcription factors. The YAP-TEAD complex triggers its target genes (CTGF and AREG) expression that promote cell proliferation and migration. Under low expression of ALDOA, cellular ATP formation is decreased due to low glycolysis. A decreased ratio of ATP/ADP leads to phosphorylation of AMPK. Knockdown of ALDOA can also trigger the formation of v-ATPase-Ragulator- AXIN/LKB1 complex, which activates AMPK in an AMP/ADP-independent manner. Activated AMPK then phosphorylates YAP, promoting its binding to 14-3-3, thus resulting in its cytoplasmic localization and functional inactivation. Moreover, phosphorylated YAP is also targeted for ubiquitylation and degradation.

Article Snippet: In this study, the primary antibodies detecting human ALDOA, AREG and CTGF (dilution 1:100; Proteintech) were used.

Techniques: Expressing, Inhibition, Binding Assay, Migration, Phospho-proteomics, Knockdown, Functional Assay

Journal: Frontiers in Oncology

Article Title: Aldolase A and Phospholipase D1 Synergistically Resist Alkylating Agents and Radiation in Lung Cancer

doi: 10.3389/fonc.2021.811635

Figure Lengend Snippet: Events and activities related to PLD depend on ALDOA. (A) The production of lysophosphatidylcholine (LPC) and lysophosphatidylethanolamine (LPE) were obtained from the available CCLE metabolomics results. Combined with previous research, we collected the survival fraction after radiation exposure in each CCLE cell. A549, H1299, H2122 (radioresistant group), and H1792, H23, H1975 (radiosensitivity group) lung cancer cells were divided into two groups to quantify their LPC and LPE concentrations. (B) The dependence of LPC or LPE products on glycolytic pathways from the available CCLE metabolites profile. These configuration files are calculated based on the RNA-seq expression of each candidate in the CCLE omics database. (C) Trends between glycolysis and phospholipase D metabolic pathways. This result shows that the performance of LPC/LPE depends on HK2, ALDOA, and PLD. The importance of TPI1 has decreased. Red color means positive dependence. Blue color means negative dependence for LPC/LPE products. (D) Two-way model of immunoprecipitation using ALDOA and PLD2 antibodies in CL1-0 cells with or without forced expression of an exogenous ALDOA gene. IgG served as the negative control. (E) The protein levels of ALDOA, PLD1, and PLD2 in A549 cells with or without shALDOA. Tubulin serves as an internal control. (F) The PLD2 protein level after treatment with MG-132 is time-dependent in A549 cells with vector control or ALDOA overexpression. Tubulin serves as an internal control. The significance of the difference was analyzed using the nonparametric Mann–Whitney U test.

Article Snippet: Polyclonal rabbit anti-human ALDOA antibody (1:100, Epitomics, Cambridge, MA, USA), anti-human PLD1 and PLD2 antibodies (1:100, GeneTex, Hsinchu, Taiwan) was used to stain slides.

Techniques: RNA Sequencing, Expressing, Immunoprecipitation, Negative Control, Control, Plasmid Preparation, Over Expression, MANN-WHITNEY

Journal: Frontiers in Oncology

Article Title: Aldolase A and Phospholipase D1 Synergistically Resist Alkylating Agents and Radiation in Lung Cancer

doi: 10.3389/fonc.2021.811635

Figure Lengend Snippet: ALDOA increases autophagy and invasion sensitivity after radiation exposure. (A) The survival fraction after radiation exposure in various lung cancer cells (CL1-0, CL1-5, H1299, and H1355). Radiation dose: 0~10 Gy. (B) The survival fraction after radiation exposure in CL1-0 cells with vector control or ALDOA overexpression. Radiation dose: 0~10 Gy. (C) Quantify the expression levels of PLD1 after radiation exposure in CL1-0 cells with vector control or ALDOA overexpression. Radiation dose: 8 Gy (D) The LC3-I/II protein level in CL1-0 cells with vector control or ALDOA overexpression. Tubulin serves as an internal control. Radiation dose: 8 Gy (E) Quantitation of the activity of caspase-3 in A549 cells after alkylating agent exposure in CL1-0 cells with vector control or ALDOA overexpression. Radiation dose: 8 Gy (F) The LC3-I/II and LAMP2 protein level after radiation exposure in a time-dependent manner in CL1-0 cells with vector control or ALDOA overexpression. Tubulin serves as an internal control. Radiation dose: 8 Gy. (G) The Ki-67, PCNA, PARP, γ-H2AX, and RRM2 protein level after radiation exposure in CL1-0 cells with vector control or ALDOA overexpression. Tubulin serves as an internal control. Radiation dose: 8 Gy. The data from three independent experiments are presented in (B–D) as the means ± SEM. The significance of the difference was analyzed using the nonparametric Mann–Whitney U test.

Article Snippet: Polyclonal rabbit anti-human ALDOA antibody (1:100, Epitomics, Cambridge, MA, USA), anti-human PLD1 and PLD2 antibodies (1:100, GeneTex, Hsinchu, Taiwan) was used to stain slides.

Techniques: Plasmid Preparation, Control, Over Expression, Expressing, Quantitation Assay, Activity Assay, MANN-WHITNEY

Journal: Frontiers in Oncology

Article Title: Aldolase A and Phospholipase D1 Synergistically Resist Alkylating Agents and Radiation in Lung Cancer

doi: 10.3389/fonc.2021.811635

Figure Lengend Snippet: ALDOA interferes with the activity of the PLD2 enzyme but increases the total PLD and phosphatidic acid products. (A) Intracellular PLD2 activity in CL1-0 cells with and without ALDOA gene overexpression. (B) Intracellular PLD2 activity in ALDOA knockdown CL1-5 cells with and without PLD1 or PLD2 overexpression. (C) Total intracellular PLD activity in an ALDOA two-way cell model. (D) Intercellular PLD2-specific activity in ALDOA-overexpressing CL1-0 cells treated with pharmaceutical PLD1 inhibitor VU0359595 and PLD2 inhibitor CAY10594, respectively. In this study, 10 μM of VU0359595 and 50 μM of CAY10594. (E) Total intracellular phosphatidic acid production in the ALDOA overexpression model. (F) Total intracellular phosphatidic acid production in CL1-0 cells overexpressing ALDOA with and without PLD1 or PLD2 gene knockdown by shRNAs. The data from three independent experiments are presented in (A–F) as the means ± SEM. The significance of the difference was analyzed using the nonparametric Mann–Whitney U test.

Article Snippet: Polyclonal rabbit anti-human ALDOA antibody (1:100, Epitomics, Cambridge, MA, USA), anti-human PLD1 and PLD2 antibodies (1:100, GeneTex, Hsinchu, Taiwan) was used to stain slides.

Techniques: Activity Assay, Over Expression, Knockdown, MANN-WHITNEY

Journal: Frontiers in Oncology

Article Title: Aldolase A and Phospholipase D1 Synergistically Resist Alkylating Agents and Radiation in Lung Cancer

doi: 10.3389/fonc.2021.811635

Figure Lengend Snippet: The prognosis value of ALDOA-PLD1 for lung cancer patients. (A) Scores (0–3) indicate PLD1 protein levels in representative lung tumor tissues. (B) The expression level of the PLD1 protein in tumor tissue compared with the corresponding normal adjacent tissue. (C) Kaplan–Meier analysis of PLD1 protein expression at concurrently low or high levels as determined by IHC staining at the endpoint of overall survival probability and disease-free survival probability in lung cancer patients. (D) Kaplan–Meier analysis of PLD1 combined with ALDOA protein expression at concurrently low or high levels or others as determined by IHC staining at the endpoint of overall survival probability in lung cancer patients. (E) Multivariate analysis of ALDOA/PLD1/PLD2 and clinical parameters in a clinical cohort. The significance of the differences in (C, D) was analyzed using the Student’s t- test.

Article Snippet: Polyclonal rabbit anti-human ALDOA antibody (1:100, Epitomics, Cambridge, MA, USA), anti-human PLD1 and PLD2 antibodies (1:100, GeneTex, Hsinchu, Taiwan) was used to stain slides.

Techniques: Expressing, Immunohistochemistry

Journal: Journal of Cellular and Molecular Medicine

Article Title: Aldolase A as a prognostic factor and mediator of progression via inducing epithelial–mesenchymal transition in gastric cancer

doi: 10.1111/jcmm.13732

Figure Lengend Snippet: Association between the expression of ALDOA and clinicopathological factors in gastric cancers

Article Snippet: Protein extracts (50 mg) were resolved on 10% sodium dodecyl sulphate–polyacrylamide gel electrophoresis gels, transferred to nitrocellulose membranes (0.45 mm), and immunoblotted with rabbit anti‐human ALDOA antibody (11217‐1‐AP; 1:1000; Proteintech), anti‐E‐cadherin antibody (ab40772; 1:1000; Abcam), anti‐N‐cadherin antibody (ab18203; 1:1000; Abcam), anti‐Vimentin (ab92547; 1:1000; Abcam) or mouse anti‐human β‐actin monoclonal antibody (ab133626; 1:1000; Abcam).

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Aldolase A as a prognostic factor and mediator of progression via inducing epithelial–mesenchymal transition in gastric cancer

doi: 10.1111/jcmm.13732

Figure Lengend Snippet: Aldolase A ( ALDOA ) was up‐regulated in gastric cancer ( GC ). A, The mRNA expression of ALDOA was found to be significantly up‐regulated in GC tissues than in adjacent normal controls in 30 patients with GC ( P < .001). B, Examples of immunohistochemical ( IHC ) staining of the expression of ALDOA in GC (I and III ) and normal gastric tissues ( II and IV ). C, The expression of ALDOA was significantly higher in GC tissues than in their adjacent normal control as determined by IHC staining (χ 2 = 8.076, P = .004)

Article Snippet: Protein extracts (50 mg) were resolved on 10% sodium dodecyl sulphate–polyacrylamide gel electrophoresis gels, transferred to nitrocellulose membranes (0.45 mm), and immunoblotted with rabbit anti‐human ALDOA antibody (11217‐1‐AP; 1:1000; Proteintech), anti‐E‐cadherin antibody (ab40772; 1:1000; Abcam), anti‐N‐cadherin antibody (ab18203; 1:1000; Abcam), anti‐Vimentin (ab92547; 1:1000; Abcam) or mouse anti‐human β‐actin monoclonal antibody (ab133626; 1:1000; Abcam).

Techniques: Expressing, Immunohistochemical staining, Immunohistochemistry, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Aldolase A as a prognostic factor and mediator of progression via inducing epithelial–mesenchymal transition in gastric cancer

doi: 10.1111/jcmm.13732

Figure Lengend Snippet: Univariate and multivariate Cox proportional hazards analyses of the expression of ALDOA gene and overall survival for patients with gastric cancer

Article Snippet: Protein extracts (50 mg) were resolved on 10% sodium dodecyl sulphate–polyacrylamide gel electrophoresis gels, transferred to nitrocellulose membranes (0.45 mm), and immunoblotted with rabbit anti‐human ALDOA antibody (11217‐1‐AP; 1:1000; Proteintech), anti‐E‐cadherin antibody (ab40772; 1:1000; Abcam), anti‐N‐cadherin antibody (ab18203; 1:1000; Abcam), anti‐Vimentin (ab92547; 1:1000; Abcam) or mouse anti‐human β‐actin monoclonal antibody (ab133626; 1:1000; Abcam).

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Aldolase A as a prognostic factor and mediator of progression via inducing epithelial–mesenchymal transition in gastric cancer

doi: 10.1111/jcmm.13732

Figure Lengend Snippet: Univariate and multivariate Cox proportional hazards analyses of the gene expression of ALDOA and disease‐free survival for patients with gastric cancer

Article Snippet: Protein extracts (50 mg) were resolved on 10% sodium dodecyl sulphate–polyacrylamide gel electrophoresis gels, transferred to nitrocellulose membranes (0.45 mm), and immunoblotted with rabbit anti‐human ALDOA antibody (11217‐1‐AP; 1:1000; Proteintech), anti‐E‐cadherin antibody (ab40772; 1:1000; Abcam), anti‐N‐cadherin antibody (ab18203; 1:1000; Abcam), anti‐Vimentin (ab92547; 1:1000; Abcam) or mouse anti‐human β‐actin monoclonal antibody (ab133626; 1:1000; Abcam).

Techniques: Gene Expression

Journal: Journal of Cellular and Molecular Medicine

Article Title: Aldolase A as a prognostic factor and mediator of progression via inducing epithelial–mesenchymal transition in gastric cancer

doi: 10.1111/jcmm.13732

Figure Lengend Snippet: Knockdown of the expression of aldolase A ( ALDOA ) suppressed the proliferation and invasion abilities of gastric cancer cells. A, Western blot and (B) RT ‐ PCR analysis were performed to determine the knockdown effect of ALDOA in AGS and MGC 803 cells. β‐Actin was used as the loading control. C, The growth curves of AGS and MGC 803 cells with transfected sh RNA – ALDOA or scramble sequence. Cell growth was determined using CCK ‐8. D, A colony formation assay was performed to determine the oncogenic growth of AGS and MGC 803 cells transfected with sh RNA or scramble sequence. E, Transwell assays were performed to determine the invasion abilities of sh RNA ‐ ALDOA ‐transfected AGS and MGC 803 cells. Values are presented as mean ± standard deviation of three independent experiments. * P < .05

Article Snippet: Protein extracts (50 mg) were resolved on 10% sodium dodecyl sulphate–polyacrylamide gel electrophoresis gels, transferred to nitrocellulose membranes (0.45 mm), and immunoblotted with rabbit anti‐human ALDOA antibody (11217‐1‐AP; 1:1000; Proteintech), anti‐E‐cadherin antibody (ab40772; 1:1000; Abcam), anti‐N‐cadherin antibody (ab18203; 1:1000; Abcam), anti‐Vimentin (ab92547; 1:1000; Abcam) or mouse anti‐human β‐actin monoclonal antibody (ab133626; 1:1000; Abcam).

Techniques: Knockdown, Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Control, Transfection, Sequencing, CCK-8 Assay, Colony Assay, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Aldolase A as a prognostic factor and mediator of progression via inducing epithelial–mesenchymal transition in gastric cancer

doi: 10.1111/jcmm.13732

Figure Lengend Snippet: Silencing the expression of aldolase A ( ALDOA ) suppressed the epithelial–mesenchymal transition in gastric cancer. Silencing the expression of ALDOA resulted in the increased expression of E‐cadherin and decreased expression of vimentin, N‐cadherin, Snail and ZEB 1, at both the protein (A) and transcriptional levels (B). C, Confocal microscopic analysis of phenotypic markers, including E‐cadherin, crystal violet and vimentin. The red signal represents the staining of corresponding protein and the blue signal represents the nuclear DNA staining using 4′,6‐diamidino‐2‐phenylindole

Article Snippet: Protein extracts (50 mg) were resolved on 10% sodium dodecyl sulphate–polyacrylamide gel electrophoresis gels, transferred to nitrocellulose membranes (0.45 mm), and immunoblotted with rabbit anti‐human ALDOA antibody (11217‐1‐AP; 1:1000; Proteintech), anti‐E‐cadherin antibody (ab40772; 1:1000; Abcam), anti‐N‐cadherin antibody (ab18203; 1:1000; Abcam), anti‐Vimentin (ab92547; 1:1000; Abcam) or mouse anti‐human β‐actin monoclonal antibody (ab133626; 1:1000; Abcam).

Techniques: Expressing, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Aldolase A as a prognostic factor and mediator of progression via inducing epithelial–mesenchymal transition in gastric cancer

doi: 10.1111/jcmm.13732

Figure Lengend Snippet: Aldolase A ( ALDOA ) affected the HIF ‐1α activity in gastric cancer. A, It could increase the HRE ‐luciferase activity in a dose‐dependent manner ( P < .05). B, Silencing the expression of ALDOA decreased the HRE activity ( P < .05)

Article Snippet: Protein extracts (50 mg) were resolved on 10% sodium dodecyl sulphate–polyacrylamide gel electrophoresis gels, transferred to nitrocellulose membranes (0.45 mm), and immunoblotted with rabbit anti‐human ALDOA antibody (11217‐1‐AP; 1:1000; Proteintech), anti‐E‐cadherin antibody (ab40772; 1:1000; Abcam), anti‐N‐cadherin antibody (ab18203; 1:1000; Abcam), anti‐Vimentin (ab92547; 1:1000; Abcam) or mouse anti‐human β‐actin monoclonal antibody (ab133626; 1:1000; Abcam).

Techniques: Activity Assay, Luciferase, Expressing