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MedChemExpress rabbit anti hkdc1
Rabbit Anti Hkdc1, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+anti+hkdc1/Phospho-Smad1+(Ser463%2FSer465)+Antibody/pm41671784-79-2-5
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rabbit anti hkdc1 - by Bioz Stars, 2026-09

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A A heatmap depicted the mRNA expression levels of glycolytic genes (as mentioned in Fig. 5E, G in high-expression and low-expression HNF4α groups within the TCGA GC cohort. The median value of HNF4α was utilized as the cutoff threshold to categorize GC patients into high-expression and low-expression HNF4α groups. B A scatter plot illustrated the correlation between HNF4α and HKDC1 mRNA expression in TCGA GC samples ( n = 408, Spearman’s correlation test). C Another scatter plot displayed the correlation between HNF4α and HKDC1 mRNA expression in CCLE GC cell lines ( n = 71, Spearman’s correlation test). D A boxplot presented HKDC1 mRNA expression in GC tissues and the corresponding normal gastric tissues in TCGA and Genotype-Tissue Expression (GTEx) databases ( n = 408 for tumor and n = 211 for normal) using GEPIA ( http://gepia.cancer-pku.cn/ ). E Assessment of the prognostic values of overall survival in GC patients based on the mRNA expression level of HKDC1 via Kaplan–Meier plotter by GEPIA (cutoff: high HKDC1 60%, low HKDC1 40%). F The expression profiles of HKDC1 across the ten identified cell types derived from the single-cell data referenced in Fig. and Supplementary Fig. were depicted. G A scatter plot illustrated the correlation between HNF4α and HKDC1 mRNA expression in single-cell RNAseq datasets, as determined by Spearman’s correlation test. H , I The expression level of HKDC1 was upregulated in HGC-27 and OCUM-1 cells by HNF4α7 overexpression and down-regulated in AGS, NUGC-4, and KATO-III cells by HNF4α knockdown, as analyzed by real-time PCR. J The expression level of HKDC1 was upregulated in HGC-27 and OCUM-1 cells by HNF4α7 overexpression and down-regulated in AGS cells by HNF4α knockdown, as analyzed by WB. Data were presented as the mean ± SEM. The difference in significance was analyzed by unpaired two-tailed Student’s t -test ( H ) or one-way ANOVA ( I ).

Journal: Cell Death & Disease

Article Title: HNF4α-HKDC1 axis orchestrates a metabolic rewiring to promote migration and metastasis in advanced gastric cancer

doi: 10.1038/s41419-026-08627-y

Figure Lengend Snippet: A A heatmap depicted the mRNA expression levels of glycolytic genes (as mentioned in Fig. 5E, G in high-expression and low-expression HNF4α groups within the TCGA GC cohort. The median value of HNF4α was utilized as the cutoff threshold to categorize GC patients into high-expression and low-expression HNF4α groups. B A scatter plot illustrated the correlation between HNF4α and HKDC1 mRNA expression in TCGA GC samples ( n = 408, Spearman’s correlation test). C Another scatter plot displayed the correlation between HNF4α and HKDC1 mRNA expression in CCLE GC cell lines ( n = 71, Spearman’s correlation test). D A boxplot presented HKDC1 mRNA expression in GC tissues and the corresponding normal gastric tissues in TCGA and Genotype-Tissue Expression (GTEx) databases ( n = 408 for tumor and n = 211 for normal) using GEPIA ( http://gepia.cancer-pku.cn/ ). E Assessment of the prognostic values of overall survival in GC patients based on the mRNA expression level of HKDC1 via Kaplan–Meier plotter by GEPIA (cutoff: high HKDC1 60%, low HKDC1 40%). F The expression profiles of HKDC1 across the ten identified cell types derived from the single-cell data referenced in Fig. and Supplementary Fig. were depicted. G A scatter plot illustrated the correlation between HNF4α and HKDC1 mRNA expression in single-cell RNAseq datasets, as determined by Spearman’s correlation test. H , I The expression level of HKDC1 was upregulated in HGC-27 and OCUM-1 cells by HNF4α7 overexpression and down-regulated in AGS, NUGC-4, and KATO-III cells by HNF4α knockdown, as analyzed by real-time PCR. J The expression level of HKDC1 was upregulated in HGC-27 and OCUM-1 cells by HNF4α7 overexpression and down-regulated in AGS cells by HNF4α knockdown, as analyzed by WB. Data were presented as the mean ± SEM. The difference in significance was analyzed by unpaired two-tailed Student’s t -test ( H ) or one-way ANOVA ( I ).

Article Snippet: Rabbit anti-HKDC1 antibody (Cat. #25874-1-AP, 1:1000 for WB) was obtained from Proteintech, Wuhan, China.

Techniques: Expressing, Derivative Assay, Single Cell, RNA sequencing, Over Expression, Knockdown, Real-time Polymerase Chain Reaction, Two Tailed Test

A – D HKDC1 knockdown significantly suppressed the peritoneal metastasis of GC cells. shCtrl and shHKDC1 sublines of AGS cells were subjected to intraperitoneal injection into nude mice (seven mice per group). Forty days later, images ( A ) and quantification ( B ) of luciferase expression, and representative images ( C ) and number ( D ) of metastatic nodules in the abdominal cavity of nude mice for each group were presented. E The diminished migratory capacity of AGS cells resulting from HNF4α knockdown was restored by HKDC1 overexpression, as demonstrated by the wound healing assay (scale bar: 100 μm). Representative images (upper panel) and quantification of wound distance (lower panel) were presented ( n = 9 biologically independent samples). F , G HKDC1 knockdown attenuated the effects of HNF4α7 overexpression on the migration of HGC-27 cells by the 3D tumor spheroid migration assay. Representative images ( F , scale bar: 50 μm) and quantification of wound distance ( G ) were shown ( n = 4 biologically independent samples). H Metabolite levels of organic acid compounds (including glycolysis/TCA cycle Intermediates) before and after HNF4α or HKDC1 knockdown in AGS cells were assayed by targeted mass spectrometry ( n = 3 biologically independent samples). I Metabolite levels of organic acid compounds (including glycolysis/TCA cycle Intermediates) of peritoneal metastases in each mouse of Fig. 6A were assayed by targeted mass spectrometry (seven mice per group). J – L The effects of HNF4α knockdown on lactate production ( J ), ROS levels ( K ), and ATP production ( L ) were restored by HKDC1 overexpression in AGS cells ( n = 3 biologically independent samples). M , N The diminished transwell migration ability of AGS cells caused by HNF4α or HKDC1 knockdown was rescued by treatment with lactate (5 mM) or α-KG (5 mM). Representative images ( M ) and quantification ( N ) were shown ( n = 3 biologically independent samples). Scale bar: 50 μm. Data were presented as the mean ± SEM. The difference significance was analyzed by unpaired two-tailed Student’s t -test ( B , D , I ) or one-way ANOVA ( E , G , H , J – L , N ).

Journal: Cell Death & Disease

Article Title: HNF4α-HKDC1 axis orchestrates a metabolic rewiring to promote migration and metastasis in advanced gastric cancer

doi: 10.1038/s41419-026-08627-y

Figure Lengend Snippet: A – D HKDC1 knockdown significantly suppressed the peritoneal metastasis of GC cells. shCtrl and shHKDC1 sublines of AGS cells were subjected to intraperitoneal injection into nude mice (seven mice per group). Forty days later, images ( A ) and quantification ( B ) of luciferase expression, and representative images ( C ) and number ( D ) of metastatic nodules in the abdominal cavity of nude mice for each group were presented. E The diminished migratory capacity of AGS cells resulting from HNF4α knockdown was restored by HKDC1 overexpression, as demonstrated by the wound healing assay (scale bar: 100 μm). Representative images (upper panel) and quantification of wound distance (lower panel) were presented ( n = 9 biologically independent samples). F , G HKDC1 knockdown attenuated the effects of HNF4α7 overexpression on the migration of HGC-27 cells by the 3D tumor spheroid migration assay. Representative images ( F , scale bar: 50 μm) and quantification of wound distance ( G ) were shown ( n = 4 biologically independent samples). H Metabolite levels of organic acid compounds (including glycolysis/TCA cycle Intermediates) before and after HNF4α or HKDC1 knockdown in AGS cells were assayed by targeted mass spectrometry ( n = 3 biologically independent samples). I Metabolite levels of organic acid compounds (including glycolysis/TCA cycle Intermediates) of peritoneal metastases in each mouse of Fig. 6A were assayed by targeted mass spectrometry (seven mice per group). J – L The effects of HNF4α knockdown on lactate production ( J ), ROS levels ( K ), and ATP production ( L ) were restored by HKDC1 overexpression in AGS cells ( n = 3 biologically independent samples). M , N The diminished transwell migration ability of AGS cells caused by HNF4α or HKDC1 knockdown was rescued by treatment with lactate (5 mM) or α-KG (5 mM). Representative images ( M ) and quantification ( N ) were shown ( n = 3 biologically independent samples). Scale bar: 50 μm. Data were presented as the mean ± SEM. The difference significance was analyzed by unpaired two-tailed Student’s t -test ( B , D , I ) or one-way ANOVA ( E , G , H , J – L , N ).

Article Snippet: Rabbit anti-HKDC1 antibody (Cat. #25874-1-AP, 1:1000 for WB) was obtained from Proteintech, Wuhan, China.

Techniques: Knockdown, Injection, Luciferase, Expressing, Over Expression, Wound Healing Assay, Migration, Mass Spectrometry, Two Tailed Test

A Overexpression of HNF4α7, but not HNF4α7(ΔDBD), resulted in an elevation of HKDC1 mRNA expression in HGC-27 and OCUM-1 cells. B Integration of HNF4α ChIP-seq data in seven HNF4α-positive GC cell lines (IM95, Ist1, KATO-III, NUGC4, OCUM-1, SNU16, and YCC3) and histone ChIP-seq data (H3K27ac, H3K4me3, and H3K4me1) in IM95 cell line from the GEO database ( GSE114018 and GSE75898 ) was performed at human HKDC1 gene locus ( NR_120648.1 ). The arrow indicated the direction of transcription. The blue frames denoted potential HNF4α binding regions, namely E1, E2, E3, E4 and E5. C HNF4α promoted the transcription of the potential HNF4α binding region E4. HEK293T cells transiently expressing or HGC-27 cells stably expressing pLV vector or pLV-HNF4α7 were cotransfected with pRL-TK and pGL6-TA constructs containing E1, E2, E3, E4, or E5, as indicated. Twenty-four hours later, cells were harvested, and a dual-luciferase reporter assay was conducted. D Two potential HNF4α response elements (HREs), namely S1 ( + 4323 ~ +4334 bp) and S2 ( + 4365 ~ +4376 bp), were predicted in the potential HNF4α binding region E4 within the human HKDC1 gene locus. The sequence (+4323 ~ +4381 bp, named S3), containing S1 and S2, was employed in subsequent experiments. E Overexpression of HNF4α7 promoted the transcription of S3, but not its mutant with mutations in both S1 and S2 mentioned in ( D ). HEK293T cells transiently expressing or HGC-27 cells stably expressing pLV vector or pLV-HNF4α7 were cotransfected with pRL-TK and pGL6-TA constructs containing S3 or its mutant, as indicated. F – H The binding of HNF4α to S3 within the human HKDC1 gene locus was detected by EMSA ( F ) and ChIP ( H ) assay. For EMSA, recombinant GST-HNF4α7 or its mutant GST-HNF4α7(ΔDBD) protein was incubated with biotin-labeled oligonucleotides of S3. Coomassie blue staining of the purified recombinant GST-HNF4α7 and GST-HNF4α7(ΔDBD) proteins is indicated in ( G ). For ChIP, HGC-27 cells (with HNF4α7 overexpressed beforehand) or AGS cells were harvested, and the ChIP assay was carried out. All data were shown as the mean ± SEM. The difference significance was analyzed by one-way ANOVA ( A ) or unpaired two-tailed Student’s t -test ( C , E ).

Journal: Cell Death & Disease

Article Title: HNF4α-HKDC1 axis orchestrates a metabolic rewiring to promote migration and metastasis in advanced gastric cancer

doi: 10.1038/s41419-026-08627-y

Figure Lengend Snippet: A Overexpression of HNF4α7, but not HNF4α7(ΔDBD), resulted in an elevation of HKDC1 mRNA expression in HGC-27 and OCUM-1 cells. B Integration of HNF4α ChIP-seq data in seven HNF4α-positive GC cell lines (IM95, Ist1, KATO-III, NUGC4, OCUM-1, SNU16, and YCC3) and histone ChIP-seq data (H3K27ac, H3K4me3, and H3K4me1) in IM95 cell line from the GEO database ( GSE114018 and GSE75898 ) was performed at human HKDC1 gene locus ( NR_120648.1 ). The arrow indicated the direction of transcription. The blue frames denoted potential HNF4α binding regions, namely E1, E2, E3, E4 and E5. C HNF4α promoted the transcription of the potential HNF4α binding region E4. HEK293T cells transiently expressing or HGC-27 cells stably expressing pLV vector or pLV-HNF4α7 were cotransfected with pRL-TK and pGL6-TA constructs containing E1, E2, E3, E4, or E5, as indicated. Twenty-four hours later, cells were harvested, and a dual-luciferase reporter assay was conducted. D Two potential HNF4α response elements (HREs), namely S1 ( + 4323 ~ +4334 bp) and S2 ( + 4365 ~ +4376 bp), were predicted in the potential HNF4α binding region E4 within the human HKDC1 gene locus. The sequence (+4323 ~ +4381 bp, named S3), containing S1 and S2, was employed in subsequent experiments. E Overexpression of HNF4α7 promoted the transcription of S3, but not its mutant with mutations in both S1 and S2 mentioned in ( D ). HEK293T cells transiently expressing or HGC-27 cells stably expressing pLV vector or pLV-HNF4α7 were cotransfected with pRL-TK and pGL6-TA constructs containing S3 or its mutant, as indicated. F – H The binding of HNF4α to S3 within the human HKDC1 gene locus was detected by EMSA ( F ) and ChIP ( H ) assay. For EMSA, recombinant GST-HNF4α7 or its mutant GST-HNF4α7(ΔDBD) protein was incubated with biotin-labeled oligonucleotides of S3. Coomassie blue staining of the purified recombinant GST-HNF4α7 and GST-HNF4α7(ΔDBD) proteins is indicated in ( G ). For ChIP, HGC-27 cells (with HNF4α7 overexpressed beforehand) or AGS cells were harvested, and the ChIP assay was carried out. All data were shown as the mean ± SEM. The difference significance was analyzed by one-way ANOVA ( A ) or unpaired two-tailed Student’s t -test ( C , E ).

Article Snippet: Rabbit anti-HKDC1 antibody (Cat. #25874-1-AP, 1:1000 for WB) was obtained from Proteintech, Wuhan, China.

Techniques: Over Expression, Expressing, ChIP-sequencing, Binding Assay, Stable Transfection, Plasmid Preparation, Construct, Luciferase, Reporter Assay, Sequencing, Mutagenesis, Recombinant, Incubation, Labeling, Staining, Purification, Two Tailed Test

A , B The suppressive effects of MMF on the mRNA expression level of HKDC1 and the migration of AGS cells by real-time PCR ( A ) and wound healing assay ( B ). Representative images ( B , upper panel) and quantification of wound distance ( B , lower panel) were presented ( n = 7 biologically independent samples). Scale bar: 100 μm. C – H MMF significantly suppressed the peritoneal metastasis of HNF4α-overexpressed HGC-27 cells, but not that of HGC-27 cells with minimal HNF4α expression in nude mice. One of the HGC-27 pLV-Vector and pLV-HNF4α7 cell sublines (all stably expressing luciferase, 1 × 10 7 cells per mouse) was intraperitoneally injected into the nude mice. Ten days later, the mice were randomly divided into two groups ( n = 7 per group), and treated with vehicle (0.9% NaCl) or MMF (40 mg/kg) once daily for 26 days by oral administration. Presented were images ( C ) and quantification ( D ) of luciferase expression, representative images ( E ) and number ( F ) of metastatic nodules in the abdominal cavity, and body weight curve ( G ) of nude mice for each group. All macroscopic metastatic lesions discernible by visual inspection were included in the quantitative analysis ( E , F ). The expression levels of HKDC1 and HNF4α in each group were examined by WB ( H ). I A proposed a working model for HNF4α-HKDC1 in the metabolism and metastasis of GC, which was antagonized by MPA. Data were presented as the mean ± SEM. The significance of differences was analyzed by one-way ANOVA ( A , D , F , G ) or unpaired two-tailed Student’s t -test ( B ).

Journal: Cell Death & Disease

Article Title: HNF4α-HKDC1 axis orchestrates a metabolic rewiring to promote migration and metastasis in advanced gastric cancer

doi: 10.1038/s41419-026-08627-y

Figure Lengend Snippet: A , B The suppressive effects of MMF on the mRNA expression level of HKDC1 and the migration of AGS cells by real-time PCR ( A ) and wound healing assay ( B ). Representative images ( B , upper panel) and quantification of wound distance ( B , lower panel) were presented ( n = 7 biologically independent samples). Scale bar: 100 μm. C – H MMF significantly suppressed the peritoneal metastasis of HNF4α-overexpressed HGC-27 cells, but not that of HGC-27 cells with minimal HNF4α expression in nude mice. One of the HGC-27 pLV-Vector and pLV-HNF4α7 cell sublines (all stably expressing luciferase, 1 × 10 7 cells per mouse) was intraperitoneally injected into the nude mice. Ten days later, the mice were randomly divided into two groups ( n = 7 per group), and treated with vehicle (0.9% NaCl) or MMF (40 mg/kg) once daily for 26 days by oral administration. Presented were images ( C ) and quantification ( D ) of luciferase expression, representative images ( E ) and number ( F ) of metastatic nodules in the abdominal cavity, and body weight curve ( G ) of nude mice for each group. All macroscopic metastatic lesions discernible by visual inspection were included in the quantitative analysis ( E , F ). The expression levels of HKDC1 and HNF4α in each group were examined by WB ( H ). I A proposed a working model for HNF4α-HKDC1 in the metabolism and metastasis of GC, which was antagonized by MPA. Data were presented as the mean ± SEM. The significance of differences was analyzed by one-way ANOVA ( A , D , F , G ) or unpaired two-tailed Student’s t -test ( B ).

Article Snippet: Rabbit anti-HKDC1 antibody (Cat. #25874-1-AP, 1:1000 for WB) was obtained from Proteintech, Wuhan, China.

Techniques: Expressing, Migration, Real-time Polymerase Chain Reaction, Wound Healing Assay, Plasmid Preparation, Stable Transfection, Luciferase, Injection, Process/Product Development, Two Tailed Test

Figure 1. Nuclear-localized HKDC1 interacts with the MLL1 complex (A) Western blot analysis of the localization of endogenous HKDC1, HK1, and HK2 in THLE3, PLC, HepG2, and Hep3B cells. Lamin B and a-tubulin served as markers for nuclear (Nuc) and cytosolic (Cyto) proteins, respectively. Ponceau staining served as a loading control.

Journal: Cell reports

Article Title: Nuclear-localized HKDC1 promotes hepatocellular carcinoma through phosphorylating RBBP5 to upregulate H3K4me3.

doi: 10.1016/j.celrep.2025.115250

Figure Lengend Snippet: Figure 1. Nuclear-localized HKDC1 interacts with the MLL1 complex (A) Western blot analysis of the localization of endogenous HKDC1, HK1, and HK2 in THLE3, PLC, HepG2, and Hep3B cells. Lamin B and a-tubulin served as markers for nuclear (Nuc) and cytosolic (Cyto) proteins, respectively. Ponceau staining served as a loading control.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit Polyclonal Anti-HKDC1 Proteintech Cat# 25874-1-AP; RRID: AB_2880279 Rabbit Polyclonal Anti-HKDC1 Abclonal Cat# E19270; RRID: AB_3675515 Rabbit Polyclonal Anti-RBBP5-pS497 Abclonal Cat# E20544; RRID: AB_3675516 Mouse Monoclonal Anti-HK1 Santa Cruz Cat# sc-46695; RRID: AB_627721 Mouse Monoclonal Anti-HK2 Santa Cruz Cat# sc-374091; RRID: AB_10917915 Rabbit Monoclonal Anti-RBBP5 CST Cat# 13171; RRID: AB_2714169 Mouse Monoclonal Anti-WDR5 Santa Cruz Cat# sc-393080; RRID: AB_3331659 Rabbit Polyclonal Anti-ASH2L Proteintech Cat# 12331-1-AP; RRID: AB_2059837 Rabbit Monoclonal Anti-MLL1c CST Cat# 14197; RRID: AB_2688010 Rabbit Monoclonal Anti-H3K4me3 CST Cat# 9751; RRID: AB_2616028 Rabbit Polyclonal Anti-H3 Proteintech Cat# 17168-1-AP; RRID: AB_2716755 Rabbit Monoclonal Anti-Thiophosphate ester Abcam Cat# ab92570; RRID: AB_10562142 Mouse Monoclonal Anti-Phosphoserine Millipore Cat# 05-1000; RRID: AB_916368 Rabbit Monoclonal Anti-ATAD5 Sangon Cat# D261469; RRID: AB_3674288 Rabbit Monoclonal Anti-CENPE Abcam Cat# ab133583; RRID: AB_2910100 Rabbit Monoclonal Anti-CDC27 Sangon Cat# D261541; RRID: AB_3674296 Rabbit Monoclonal Anti-KIF14 Sangon Cat# D163049; RRID: AB_3674320 Rabbit Polyclonal Anti-KNL1 Abclonal Cat# A13108; RRID: AB_2759957 Rabbit Monoclonal Anti-NEK7 CST Cat# 3057; RRID: AB_2150676 Mouse Monoclonal Anti-Flag-M2 Sigma Cat# F1804; RRID: AB_262044 Monoclonal Anti-HA-tag-HRP-conjugate CST Cat# 2999; RRID: AB_1264166 Rabbit Polyclonal Anti-GST-tag Proteintech Cat# 10000-0-AP; RRID: AB_11042316 Mouse Monoclonal Anti-His-tag Proteintech Cat# 66005-1-Ig; RRID: AB_11232599 Mouse Monoclonal Anti-b-Actin Proteintech Cat# 66009-1-Ig; RRID: AB_2687938 Rabbit Polyclonal Anti-LaminB1 Proteintech Cat# 12987-1-AP; RRID: AB_2136290 Mouse Monoclonal Anti-a-Tubulin Proteintech Cat# 66031-1-Ig; RRID: AB_11042766 Rabbit Polyclonal Anti-Calnexin Proteintech Cat# 10427-2-AP; RRID: AB_2069033 Bacterial and virus strains Bacteria: DH5a Tsingke Cat# TSC-C01 Bacteria: Rosetta (DE3) Tsingke Cat# TSC-E04 Biological samples Paired HCC and noncancerous tissues The First Affiliated Hospital of USTC N/A Chemicals, peptides, and recombinant proteins Kinase inhibitor library Topscience Cat# L1600 Dinaciclib Topscience Cat# T1912 OICR-9429 Topscience Cat# T6916 Polybrene Sigma Cat# H9268 Puromycin Sigma Cat# P8833 PEI Polysciences Cat# 23966-2 Penicillin-streptomycin HyClone Cat# SV30010 Certified Fetal Bovine Serum VivaCell Cat# C04001-500 (Continued on next page) Cell Reports 44, 115250, February 25, 2025 15

Techniques: Western Blot, Staining, Control

Figure 2. HKDC1 phosphorylates RBBP5 at Ser497 and promotes MLL1 complex assembly (A) Bacterially puriﬁed His-RBBP5 was incubated with or without His-HKDC1 proteins in the presence of ATP-g-S for an in vitro kinase assay. (B) Bacterially puriﬁed His-tagged WT RBBP5, the S350A mutant, or the S497A mutant was incubated with or without His-HKDC1 proteins in the presence of ATP-g-S for an in vitro kinase assay. (legend continued on next page)

Journal: Cell reports

Article Title: Nuclear-localized HKDC1 promotes hepatocellular carcinoma through phosphorylating RBBP5 to upregulate H3K4me3.

doi: 10.1016/j.celrep.2025.115250

Figure Lengend Snippet: Figure 2. HKDC1 phosphorylates RBBP5 at Ser497 and promotes MLL1 complex assembly (A) Bacterially puriﬁed His-RBBP5 was incubated with or without His-HKDC1 proteins in the presence of ATP-g-S for an in vitro kinase assay. (B) Bacterially puriﬁed His-tagged WT RBBP5, the S350A mutant, or the S497A mutant was incubated with or without His-HKDC1 proteins in the presence of ATP-g-S for an in vitro kinase assay. (legend continued on next page)

Techniques: Incubation, In Vitro, Kinase Assay, Mutagenesis

Figure 3. HKDC1-mediated phosphorylation of RBBP5 upregulates H3K4me3 to increase the expression of mitosis-related genes (A) Western blot analysis of the indicated histone methylation markers in Hep3B cells infected with viruses expressing NTC, sh-HKDC1#1, or sh-HKDC1#2 and Hep3B cells overexpressing WT HKDC1 or the KD mutant following knockdown of endogenous HKDC1 via sh-HKDC1#1. (B) Western blot analysis of the H3K4me3 level in Hep3B cells infected with viruses expressing NTC, sh-RBBP5#1, or sh-RBBP5#2 following overexpression of HKDC1. (legend continued on next page)

Journal: Cell reports

Article Title: Nuclear-localized HKDC1 promotes hepatocellular carcinoma through phosphorylating RBBP5 to upregulate H3K4me3.

doi: 10.1016/j.celrep.2025.115250

Figure Lengend Snippet: Figure 3. HKDC1-mediated phosphorylation of RBBP5 upregulates H3K4me3 to increase the expression of mitosis-related genes (A) Western blot analysis of the indicated histone methylation markers in Hep3B cells infected with viruses expressing NTC, sh-HKDC1#1, or sh-HKDC1#2 and Hep3B cells overexpressing WT HKDC1 or the KD mutant following knockdown of endogenous HKDC1 via sh-HKDC1#1. (B) Western blot analysis of the H3K4me3 level in Hep3B cells infected with viruses expressing NTC, sh-RBBP5#1, or sh-RBBP5#2 following overexpression of HKDC1. (legend continued on next page)

Techniques: Phospho-proteomics, Expressing, Western Blot, Methylation, Infection, Mutagenesis, Knockdown, Over Expression

Figure 5. HKDC1-mediated phosphorylation of RBBP5 correlates with poor prognosis in patients with HCC (A) Western blot analysis of HKDC1 and pS497 levels in paired adjacent noncancerous liver tissues (N) and human HCC tissues (T). Calnexin served as a loading control (n = 12 patients with HCC).

Journal: Cell reports

Article Title: Nuclear-localized HKDC1 promotes hepatocellular carcinoma through phosphorylating RBBP5 to upregulate H3K4me3.

doi: 10.1016/j.celrep.2025.115250

Figure Lengend Snippet: Figure 5. HKDC1-mediated phosphorylation of RBBP5 correlates with poor prognosis in patients with HCC (A) Western blot analysis of HKDC1 and pS497 levels in paired adjacent noncancerous liver tissues (N) and human HCC tissues (T). Calnexin served as a loading control (n = 12 patients with HCC).

Techniques: Phospho-proteomics, Western Blot, Control

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