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rabbit anti-human/mouse hk2 (Cell Signaling Technology Inc)

Name: rabbit anti-human/mouse hk2
Brand: Cell Signaling Technology Inc
Rating: 90 (1 reviews)

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Cell Signaling Technology Inc rabbit anti-human/mouse hk2
Rabbit Anti Human/Mouse Hk2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit anti-human/mouse hk2/product/Cell Signaling Technology Inc
Average 90 stars, based on 1 article reviews

rabbit anti-human/mouse hk2 - by Bioz Stars, 2026-02

90/100 stars

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HK2 translocates to mitochondria after CoCl 2 and PDGF stimulation. (A) Confocal microscopy of RA FLS (n = 3) after stimulation with 150 µM cobalt chloride (CoCl 2 ) for 30 and 60 min with quantification. HK2 protein is stained with green fluorescence, ATP5a is stained with red fluorescence, and DAPI is stained with blue fluorescence as control. Overlapping yellow color indicates HK2 colocalization to mitochondria. n = 3 independent experiments. The comparison between groups was performed using unpaired two-tailed Student’s t -test. Statistical significance was considered when p-value ≤ 0.05. (B) Confocal microscopy of RA FLS (n = 3) after stimulation of PDGF (20 ng/ml) after 30 and 60 min with quantification. HK2 protein is stained with green fluorescence and ATP5a is stained with red fluorescence. Overlapping yellow color indicates HK2 colocalization to mitochondria. n = 2 independent experiments. The comparison between groups was performed using unpaired two-tailed student t -test. Statistical significance was considered when p-value ≤ 0.05. *P ≤ 0.05.

Journal: Frontiers in Immunology

Article Title: Role of mitochondria-bound HK2 in rheumatoid arthritis fibroblast-like synoviocytes

doi: 10.3389/fimmu.2023.1103231

Figure Lengend Snippet: HK2 translocates to mitochondria after CoCl 2 and PDGF stimulation. (A) Confocal microscopy of RA FLS (n = 3) after stimulation with 150 µM cobalt chloride (CoCl 2 ) for 30 and 60 min with quantification. HK2 protein is stained with green fluorescence, ATP5a is stained with red fluorescence, and DAPI is stained with blue fluorescence as control. Overlapping yellow color indicates HK2 colocalization to mitochondria. n = 3 independent experiments. The comparison between groups was performed using unpaired two-tailed Student’s t -test. Statistical significance was considered when p-value ≤ 0.05. (B) Confocal microscopy of RA FLS (n = 3) after stimulation of PDGF (20 ng/ml) after 30 and 60 min with quantification. HK2 protein is stained with green fluorescence and ATP5a is stained with red fluorescence. Overlapping yellow color indicates HK2 colocalization to mitochondria. n = 2 independent experiments. The comparison between groups was performed using unpaired two-tailed student t -test. Statistical significance was considered when p-value ≤ 0.05. *P ≤ 0.05.

Article Snippet: After blocking for 1 h with 5% milk, blots were incubated overnight at 4 ° C with the following antibodies: rabbit anti-human/mouse HK2 at 1:500 (#2867, C64G5, Cell Signaling Technology, Danvers, MA), mouse anti-human HK2 at 1:500 (sc-374091, B8, Santa Cruz Co, Santa Clara, CA), mouse anti-human tubulin at 1:1,000 (Santa Cruz Co., Santa Clara, CA), rabbit anti-TOM20 at 1:500 (Cell Signaling Technology, Danvers, MA), and Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) and tubulin (Santa Cruz Co., Santa Clara, CA).

Techniques: Confocal Microscopy, Staining, Fluorescence, Control, Comparison, Two Tailed Test

Effect of mitochondrial mutant HK2 on RA FLS invasion and migration. (A, B) RA FLSs were plated in matrigel spheroids as described in methods after adenovirus infection with Ad-GFP control, Ad-HK2FL, and Ad- HK2ΔN constructs for 2 days. (A) Analysis of area of invasion after infection with adenovirus. The comparison between groups was performed using ordinary one-way ANOVA followed by Tukey’s multiple comparisons test. Statistical analysis of P-value ≤ 0.05 was considered significant. (B) Representative images of invasion with the adenovirus constructs with and without PDGF. (C, D) RA FLSs were plated in six-well plates and scratched with the end of a sterile pipette tip after infection with adenovirus constructs. Cells were left to migrate 24 h with and without PDGF (10 ng/ml) stimulation. (C) Quantification of scratch length. The comparison between groups was performed using Kruskal–Wallis test followed by Dunn’s multiple comparisons test. Statistical significance was considered when p-value ≤ 0.05. (D) Representative images of migration with different adenovirus constructs with and without PDGF stimulation. ****P ≤ 0.001; **P ≤ 0.01.

Journal: Frontiers in Immunology

Article Title: Role of mitochondria-bound HK2 in rheumatoid arthritis fibroblast-like synoviocytes

doi: 10.3389/fimmu.2023.1103231

Figure Lengend Snippet: Effect of mitochondrial mutant HK2 on RA FLS invasion and migration. (A, B) RA FLSs were plated in matrigel spheroids as described in methods after adenovirus infection with Ad-GFP control, Ad-HK2FL, and Ad- HK2ΔN constructs for 2 days. (A) Analysis of area of invasion after infection with adenovirus. The comparison between groups was performed using ordinary one-way ANOVA followed by Tukey’s multiple comparisons test. Statistical analysis of P-value ≤ 0.05 was considered significant. (B) Representative images of invasion with the adenovirus constructs with and without PDGF. (C, D) RA FLSs were plated in six-well plates and scratched with the end of a sterile pipette tip after infection with adenovirus constructs. Cells were left to migrate 24 h with and without PDGF (10 ng/ml) stimulation. (C) Quantification of scratch length. The comparison between groups was performed using Kruskal–Wallis test followed by Dunn’s multiple comparisons test. Statistical significance was considered when p-value ≤ 0.05. (D) Representative images of migration with different adenovirus constructs with and without PDGF stimulation. ****P ≤ 0.001; **P ≤ 0.01. "ns" denotes "not significant".

Techniques: Mutagenesis, Migration, Infection, Control, Construct, Comparison, Sterility, Transferring

Treatment with methyl jasmonate (MJ), which dissociates HK2 from mitochondria, impaired FLS invasion and migration. (A, B) RA FLSs (n = 4) were plated in matrigel spheroids and given vehicle (ethanol) or MJ 1 h before PDGF stimulation. (A) Quantification of area of invasion after MJ treatment. Comparison between groups was performed using Kruskal–Wallis test followed by Dunn’s multiple comparisons test. Statistical analysis of P-value ≤ 0.05 was considered significant. (B) Representative images of invasion with MJ. (C, D) RA FLSs were plated in six-well plates and scratched with the end of a sterile pipette tip. Cells were given vehicle (ethanol) or MJ 1 h before PDGF stimulation and allowed to migrate for 24 h, (C) Quantification of scratch length. The comparison between groups was performed using ordinary one-way ANOVA followed by Tukey’s multiple comparisons test. Statistical analysis of P-value ≤ 0.05 was considered significant. (D) Representative images of MJ migration. (E, F) RA FLSs were plated in 12-well plates and treated with vehicle (ethanol) or MJ 1 h before addition of 0 µM H 2 O 2 , 75 µM H 2 O 2 , and 200 µM H 2 O 2 to assess cell death and were fixed and stained after 4 h, (E) Analysis of mean gray value of cells using Fiji software. Comparison between groups was performed using Kruskal–Wallis test followed by Dunn’s multiple comparisons test. Statistical analysis of P-value ≤ 0.05 was considered significant. (F) Representative images of control and 1.5 mM MJ. *P ≤ 0.05.

Journal: Frontiers in Immunology

Article Title: Role of mitochondria-bound HK2 in rheumatoid arthritis fibroblast-like synoviocytes

doi: 10.3389/fimmu.2023.1103231

Figure Lengend Snippet: Treatment with methyl jasmonate (MJ), which dissociates HK2 from mitochondria, impaired FLS invasion and migration. (A, B) RA FLSs (n = 4) were plated in matrigel spheroids and given vehicle (ethanol) or MJ 1 h before PDGF stimulation. (A) Quantification of area of invasion after MJ treatment. Comparison between groups was performed using Kruskal–Wallis test followed by Dunn’s multiple comparisons test. Statistical analysis of P-value ≤ 0.05 was considered significant. (B) Representative images of invasion with MJ. (C, D) RA FLSs were plated in six-well plates and scratched with the end of a sterile pipette tip. Cells were given vehicle (ethanol) or MJ 1 h before PDGF stimulation and allowed to migrate for 24 h, (C) Quantification of scratch length. The comparison between groups was performed using ordinary one-way ANOVA followed by Tukey’s multiple comparisons test. Statistical analysis of P-value ≤ 0.05 was considered significant. (D) Representative images of MJ migration. (E, F) RA FLSs were plated in 12-well plates and treated with vehicle (ethanol) or MJ 1 h before addition of 0 µM H 2 O 2 , 75 µM H 2 O 2 , and 200 µM H 2 O 2 to assess cell death and were fixed and stained after 4 h, (E) Analysis of mean gray value of cells using Fiji software. Comparison between groups was performed using Kruskal–Wallis test followed by Dunn’s multiple comparisons test. Statistical analysis of P-value ≤ 0.05 was considered significant. (F) Representative images of control and 1.5 mM MJ. *P ≤ 0.05. "ns" denotes "not significant".

Techniques: Migration, Comparison, Sterility, Transferring, Staining, Software, Control

Effect of common RA drugs on mitochondrial translocation of HK2. Confocal microscopy of RA FLS (n = 3) after 60 min of stimulation of 150 µM CoCl 2 with and without either 1 µM tofacitinib for 60 min or 1 µM MTX overnight with quantification. HK2 protein is stained with green fluorescence, and ATP5a is stained with red fluorescence. Overlapping yellow color indicates HK2 colocalization to mitochondria. n = 3 independent experiments. The comparison between groups was performed using ordinary one-way ANOVA followed by Tukey’s multiple comparisons test. Statistical analysis of P-value ≤ 0.05 was considered significant. *P ≤ 0.05.

Journal: Frontiers in Immunology

Article Title: Role of mitochondria-bound HK2 in rheumatoid arthritis fibroblast-like synoviocytes

doi: 10.3389/fimmu.2023.1103231

Figure Lengend Snippet: Effect of common RA drugs on mitochondrial translocation of HK2. Confocal microscopy of RA FLS (n = 3) after 60 min of stimulation of 150 µM CoCl 2 with and without either 1 µM tofacitinib for 60 min or 1 µM MTX overnight with quantification. HK2 protein is stained with green fluorescence, and ATP5a is stained with red fluorescence. Overlapping yellow color indicates HK2 colocalization to mitochondria. n = 3 independent experiments. The comparison between groups was performed using ordinary one-way ANOVA followed by Tukey’s multiple comparisons test. Statistical analysis of P-value ≤ 0.05 was considered significant. *P ≤ 0.05.

Techniques: Translocation Assay, Confocal Microscopy, Staining, Fluorescence, Comparison

MJ decreased the expression of genes associated to HK2 identified by scRNA-seq. (A) UMAP of scRNAseq data of live CD45 − cells from digested hind limbs with main cell labels (n = 3 mice). Vasc, vascular cells; Peri. Vasc, perivascular; Contam., contamination; Osteo., osteoblasts; Chondr., chondrocytes. (B) Fibroblast subsets with cell labels based on reads of Hk2 gene. (C) Gene expression (GEX) of top 10 marker genes in Hk2-positive cells compared with Hk2 -egative cells with GO term. (D) Heatmap of differential expressed genes between disease model time point (rest, resolved, peak, and resolving) in Hk2 -positive cells. (E) Volcano plots of differentially expressed genes between the indicated conditional in Hk2 -positive cells. Each dot represents a gene. Genes in red: adjusted p-value < 0.05 and fold change > 0.5 and < −0.5. P-value calculated using FindMarkers() (Seurat) and the Wilcox method. (F) qPCR analysis of the indicated genes in RA FLS 6 h of MJ treatment. Results are average of three different RA FLS lines. Statistical analysis of P-value ≤ 0.05 was considered significant. *P ≤ 0.05 ; **P ≤ 0.01; ***P ≤ 0.001.

Journal: Frontiers in Immunology

Article Title: Role of mitochondria-bound HK2 in rheumatoid arthritis fibroblast-like synoviocytes

doi: 10.3389/fimmu.2023.1103231

Figure Lengend Snippet: MJ decreased the expression of genes associated to HK2 identified by scRNA-seq. (A) UMAP of scRNAseq data of live CD45 − cells from digested hind limbs with main cell labels (n = 3 mice). Vasc, vascular cells; Peri. Vasc, perivascular; Contam., contamination; Osteo., osteoblasts; Chondr., chondrocytes. (B) Fibroblast subsets with cell labels based on reads of Hk2 gene. (C) Gene expression (GEX) of top 10 marker genes in Hk2-positive cells compared with Hk2 -egative cells with GO term. (D) Heatmap of differential expressed genes between disease model time point (rest, resolved, peak, and resolving) in Hk2 -positive cells. (E) Volcano plots of differentially expressed genes between the indicated conditional in Hk2 -positive cells. Each dot represents a gene. Genes in red: adjusted p-value < 0.05 and fold change > 0.5 and < −0.5. P-value calculated using FindMarkers() (Seurat) and the Wilcox method. (F) qPCR analysis of the indicated genes in RA FLS 6 h of MJ treatment. Results are average of three different RA FLS lines. Statistical analysis of P-value ≤ 0.05 was considered significant. *P ≤ 0.05 ; **P ≤ 0.01; ***P ≤ 0.001.

Techniques: Expressing, Gene Expression, Marker

Adenovirus injection and Ad-15G treatment in AIA model. (A, B) B6 mice (n = 10) injected with AdenoHK2FL or Adeno HK2ΔN in either knee as described in methods. (A) Histological scores of mouse knees measuring synovial infiltration and infiltration of either HK2FL or HK2ΔN. The comparison between groups was performed using two-tailed Mann–Whitney non-parametric test for each score type. Statistical significance was considered when p-value ≤ 0.05. (B) Representative images of knee joints of mice. Asterisk (*) shows synovium (C, D) AIA mouse model (n = 10) with treatment of 15G peptide that dissociates HK2 from mitochondria as described in methods. (C) Histological score of mouse knees measuring synovial infiltration or infiltration of mice injected with Ad-GFP control or Ad-15G. The comparison between groups was performed using two-tailed Mann–Whitney non-parametric test for each score type. Statistical significance was considered when p-value ≤ 0.05. (D) Representative images of mouse joints.

Journal: Frontiers in Immunology

Article Title: Role of mitochondria-bound HK2 in rheumatoid arthritis fibroblast-like synoviocytes

doi: 10.3389/fimmu.2023.1103231

Figure Lengend Snippet: Adenovirus injection and Ad-15G treatment in AIA model. (A, B) B6 mice (n = 10) injected with AdenoHK2FL or Adeno HK2ΔN in either knee as described in methods. (A) Histological scores of mouse knees measuring synovial infiltration and infiltration of either HK2FL or HK2ΔN. The comparison between groups was performed using two-tailed Mann–Whitney non-parametric test for each score type. Statistical significance was considered when p-value ≤ 0.05. (B) Representative images of knee joints of mice. Asterisk (*) shows synovium (C, D) AIA mouse model (n = 10) with treatment of 15G peptide that dissociates HK2 from mitochondria as described in methods. (C) Histological score of mouse knees measuring synovial infiltration or infiltration of mice injected with Ad-GFP control or Ad-15G. The comparison between groups was performed using two-tailed Mann–Whitney non-parametric test for each score type. Statistical significance was considered when p-value ≤ 0.05. (D) Representative images of mouse joints.

Techniques: Injection, Comparison, Two Tailed Test, MANN-WHITNEY, Control

Journal: Cell host & microbe

Article Title: Legionella-infected macrophages engage the alveolar epithelium to metabolically reprogram myeloid cells and promote antibacterial inflammation

doi: 10.1016/j.chom.2020.07.019

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit anti-Mouse/Human HK2 (clone C64G5) , Cell Signaling Technology , Cat#2867; RRID: AB_2232946.

Techniques: Virus, Recombinant, Reverse Transcription, Membrane, Isolation, Enzyme-linked Immunosorbent Assay, Plex Assay, Flow Cytometry, Software

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