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mouse anti jmjd6 antibody  (Santa Cruz Biotechnology)
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Santa Cruz Biotechnology mouse anti jmjd6 antibody
Mouse Anti Jmjd6 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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jmjd6 antibody  (Santa Cruz Biotechnology)
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Santa Cruz Biotechnology jmjd6 antibody
Fig. 4 <t>JMJD6</t> functions as a negative modulator in regulating lung cancer cell ferroptosis. (A) Volcano plot of different gene expression profiles in H1299 cells overexpressing JMJD6. The genes marked in gray were not significantly different, while the genes marked in red and blue showed significant changes in expression levels. (B) Heatmap shows the expression ratio of significantly differentially expressed genes (fold change > 1, P < 0.05) in RNA-seq analysis. (C) Gene Ontology (GO) was used for functional annotation and enrichment analysis of up-regulated and down-regulated genes. Among the up- regulated genes, the top three in BP, CC and MF were selected for enrichment analysis. Among down-regulated genes, the top three enrichment in BP, CC were selected for analysis, while the top two enrichment in MF were selected for analysis. (D) The up-regulated and down-regulated Genes were analyzed by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. (E) Flow cytometry analysis of ROS in JMJD6 overexpression H460 cells. Representative images are shown on the left and statistical data analysis on the right. (F) The changes of lipid peroxidation level in JMJD6 over-expressed H1299 cells were detected by MDA method. (G) JC-1 fluorescent probe was used to detect the degree of mitochondrial membrane potential depolarization in H1299 cells with JMJD6 overexpression. Representative images are shown on the left and statistical data analysis on the right. The scale bars represent 50 μm. (H) ROS content in JMJD6 knockdown H157 cells was detected by flow cytometry. Representative images are shown on the left and statistical data analysis on the right. (I) MDA assay was used to detect the changes of lipid peroxidation in JMJD6 knockdown A549 cells. (J) JC-1 fluorescent probe was used to detect the changes of mitochondrial membrane potential in JMJD6 knockdown A549 cells. Representative images are shown on the left and statistical data analysis on the right. The scale bars represent 50 μm. (K) The JMJD6 knockdown A549 cells were treated with erastin and fer-1, respectively, and the cell viability was detected. (L, M, N) Tumor images of nude mice in control group, overexpressed JMJD6 group and overexpressed JMJD6 with injected 30 mg/kg erastin group (L). Tumor weight (M) and growth curve (N) were measured. Data are presented as mean ± standard deviation. Data are presented as mean ± standard deviation. *p < 0.05; **p < 0.01; ***p < 0.001
Jmjd6 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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protein  (Santa Cruz Biotechnology)
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Santa Cruz Biotechnology protein
Fig. 4 <t>JMJD6</t> functions as a negative modulator in regulating lung cancer cell ferroptosis. (A) Volcano plot of different gene expression profiles in H1299 cells overexpressing JMJD6. The genes marked in gray were not significantly different, while the genes marked in red and blue showed significant changes in expression levels. (B) Heatmap shows the expression ratio of significantly differentially expressed genes (fold change > 1, P < 0.05) in RNA-seq analysis. (C) Gene Ontology (GO) was used for functional annotation and enrichment analysis of up-regulated and down-regulated genes. Among the up- regulated genes, the top three in BP, CC and MF were selected for enrichment analysis. Among down-regulated genes, the top three enrichment in BP, CC were selected for analysis, while the top two enrichment in MF were selected for analysis. (D) The up-regulated and down-regulated Genes were analyzed by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. (E) Flow cytometry analysis of ROS in JMJD6 overexpression H460 cells. Representative images are shown on the left and statistical data analysis on the right. (F) The changes of lipid peroxidation level in JMJD6 over-expressed H1299 cells were detected by MDA method. (G) JC-1 fluorescent probe was used to detect the degree of mitochondrial membrane potential depolarization in H1299 cells with JMJD6 overexpression. Representative images are shown on the left and statistical data analysis on the right. The scale bars represent 50 μm. (H) ROS content in JMJD6 knockdown H157 cells was detected by flow cytometry. Representative images are shown on the left and statistical data analysis on the right. (I) MDA assay was used to detect the changes of lipid peroxidation in JMJD6 knockdown A549 cells. (J) JC-1 fluorescent probe was used to detect the changes of mitochondrial membrane potential in JMJD6 knockdown A549 cells. Representative images are shown on the left and statistical data analysis on the right. The scale bars represent 50 μm. (K) The JMJD6 knockdown A549 cells were treated with erastin and fer-1, respectively, and the cell viability was detected. (L, M, N) Tumor images of nude mice in control group, overexpressed JMJD6 group and overexpressed JMJD6 with injected 30 mg/kg erastin group (L). Tumor weight (M) and growth curve (N) were measured. Data are presented as mean ± standard deviation. Data are presented as mean ± standard deviation. *p < 0.05; **p < 0.01; ***p < 0.001
Protein, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti jmjd6  (Santa Cruz Biotechnology)
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Santa Cruz Biotechnology anti jmjd6

Anti Jmjd6, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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sc 28348  (Santa Cruz Biotechnology)
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Santa Cruz Biotechnology sc 28348

Sc 28348, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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jmjd6  (Santa Cruz Biotechnology)
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Santa Cruz Biotechnology jmjd6
Figure 2. <t>JMJD6</t> is required for neuroblastoma growth and facilitates MYC-mediated cellular transformation. (A) Copy number of genes encoding JmjC domain proteins in St Jude neuroblastoma cohort (https://platform.stjude.cloud). (B) Kaplan-Meier survival curve showing high JMJD6 is correlated with worse event-free survival (SEQC RNA-seq dataset). (C) Crystal violet showing the colony staining on day 7 after JMJD6 shRNA knockdown in neuroblastoma cell lines validated by western blot (harvested at 72 hr). n=single experiment. (D) Xenograft tumor growth of BE2C (right) models with lentiviral JMJD6 shRNA knockdown. p-Value calculated by multiple unpaired t-test across each row. n=5 per group. ***p<0.001, **p<0.01. (E) Xenograft tumor growth of SK-N-AS models with lentiviral JMJD6 shRNA knockdown. p-value calculated by multiple unpaired t-test across each row. ***p<0.001, **p<0.01. (F) Western blot validating the expression of retroviral-based MYCN and JMJD6 in JoMa1 cells. (G) Cell proliferation of JoMa1 cells transduced with indicated constructs expressing GFP, JMJD6, MYCN, JMJD6+MYCN. (H) Colony formation of JoMa1 cells transduced with indicated constructs, GFP, JMJD6, MYCN, JMJD6+MYCN. Top panel showing photos taken under light microscope. Bottom panel showing cell colonies stained with crystal violet. (I) Xenograft tumor growth of JoMa1 cells transduced with indicated constructs, GFP, JMJD6, MYCN, JMJD6+MYCN. n=5 per group. p-Value calculated by multiple unpaired t-test across each row. ***p<0.001, **p<0.01. Data are Mean ± SEM.
Jmjd6, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti jmjd6 antibody  (Santa Cruz Biotechnology)
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Santa Cruz Biotechnology anti jmjd6 antibody
Figure 2. <t>JMJD6</t> is required for neuroblastoma growth and facilitates MYC-mediated cellular transformation. (A) Copy number of genes encoding JmjC domain proteins in St Jude neuroblastoma cohort (https://platform.stjude.cloud). (B) Kaplan-Meier survival curve showing high JMJD6 is correlated with worse event-free survival (SEQC RNA-seq dataset). (C) Crystal violet showing the colony staining on day 7 after JMJD6 shRNA knockdown in neuroblastoma cell lines validated by western blot (harvested at 72 hr). n=single experiment. (D) Xenograft tumor growth of BE2C (right) models with lentiviral JMJD6 shRNA knockdown. p-Value calculated by multiple unpaired t-test across each row. n=5 per group. ***p<0.001, **p<0.01. (E) Xenograft tumor growth of SK-N-AS models with lentiviral JMJD6 shRNA knockdown. p-value calculated by multiple unpaired t-test across each row. ***p<0.001, **p<0.01. (F) Western blot validating the expression of retroviral-based MYCN and JMJD6 in JoMa1 cells. (G) Cell proliferation of JoMa1 cells transduced with indicated constructs expressing GFP, JMJD6, MYCN, JMJD6+MYCN. (H) Colony formation of JoMa1 cells transduced with indicated constructs, GFP, JMJD6, MYCN, JMJD6+MYCN. Top panel showing photos taken under light microscope. Bottom panel showing cell colonies stained with crystal violet. (I) Xenograft tumor growth of JoMa1 cells transduced with indicated constructs, GFP, JMJD6, MYCN, JMJD6+MYCN. n=5 per group. p-Value calculated by multiple unpaired t-test across each row. ***p<0.001, **p<0.01. Data are Mean ± SEM.
Anti Jmjd6 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti jmjd6 antibody/product/Santa Cruz Biotechnology
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anti jmjd6 sc 28348 human  (Santa Cruz Biotechnology)
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Santa Cruz Biotechnology anti jmjd6 sc 28348 human
Fig. 4 Tumor-CM activates THP-1 cells and upregulates <t>JMJD6</t> expression. a Morphology of THP-1 cells cultured in normal medium, with or without PMA and tumor-CM (H460) for 48 h. Scale bar, 100 μm. b The changes of macrophage related markers on the surface of THP-1 cells were detected by flow cytometry after PMA induction. Representative results. c CD206 expression of TAM surfaces were detected by flow cytometry after tumor-CM induction. d The secretion of IL-10 in the TAM culture supernatant detected by CBA assays. e THP-1 cells were treated with PMA alone or tumor-CM for 2 or 3 days, followed by JMJD6 protein level detection. f Representative images of immunofluorescence staining of CD68 (red), JMJD6 (green) and DAPI (blue) in THP-1 cells treated with PMA alone or tumor-CM for 2 days, the scale bars 10 μm and 20 μm (right panel) as indicated in figures. Data are shown as mean ± SD (n = 3). *P < 0.05, **P < 0.01.
Anti Jmjd6 Sc 28348 Human, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Fig. 4 JMJD6 functions as a negative modulator in regulating lung cancer cell ferroptosis. (A) Volcano plot of different gene expression profiles in H1299 cells overexpressing JMJD6. The genes marked in gray were not significantly different, while the genes marked in red and blue showed significant changes in expression levels. (B) Heatmap shows the expression ratio of significantly differentially expressed genes (fold change > 1, P < 0.05) in RNA-seq analysis. (C) Gene Ontology (GO) was used for functional annotation and enrichment analysis of up-regulated and down-regulated genes. Among the up- regulated genes, the top three in BP, CC and MF were selected for enrichment analysis. Among down-regulated genes, the top three enrichment in BP, CC were selected for analysis, while the top two enrichment in MF were selected for analysis. (D) The up-regulated and down-regulated Genes were analyzed by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. (E) Flow cytometry analysis of ROS in JMJD6 overexpression H460 cells. Representative images are shown on the left and statistical data analysis on the right. (F) The changes of lipid peroxidation level in JMJD6 over-expressed H1299 cells were detected by MDA method. (G) JC-1 fluorescent probe was used to detect the degree of mitochondrial membrane potential depolarization in H1299 cells with JMJD6 overexpression. Representative images are shown on the left and statistical data analysis on the right. The scale bars represent 50 μm. (H) ROS content in JMJD6 knockdown H157 cells was detected by flow cytometry. Representative images are shown on the left and statistical data analysis on the right. (I) MDA assay was used to detect the changes of lipid peroxidation in JMJD6 knockdown A549 cells. (J) JC-1 fluorescent probe was used to detect the changes of mitochondrial membrane potential in JMJD6 knockdown A549 cells. Representative images are shown on the left and statistical data analysis on the right. The scale bars represent 50 μm. (K) The JMJD6 knockdown A549 cells were treated with erastin and fer-1, respectively, and the cell viability was detected. (L, M, N) Tumor images of nude mice in control group, overexpressed JMJD6 group and overexpressed JMJD6 with injected 30 mg/kg erastin group (L). Tumor weight (M) and growth curve (N) were measured. Data are presented as mean ± standard deviation. Data are presented as mean ± standard deviation. *p < 0.05; **p < 0.01; ***p < 0.001

Journal: Journal of translational medicine

Article Title: JMJD6 K375 acetylation restrains lung cancer progression by enhancing METTL14/m6A/SLC3A2 axis mediated cell ferroptosis.

doi: 10.1186/s12967-025-06241-8

Figure Lengend Snippet: Fig. 4 JMJD6 functions as a negative modulator in regulating lung cancer cell ferroptosis. (A) Volcano plot of different gene expression profiles in H1299 cells overexpressing JMJD6. The genes marked in gray were not significantly different, while the genes marked in red and blue showed significant changes in expression levels. (B) Heatmap shows the expression ratio of significantly differentially expressed genes (fold change > 1, P < 0.05) in RNA-seq analysis. (C) Gene Ontology (GO) was used for functional annotation and enrichment analysis of up-regulated and down-regulated genes. Among the up- regulated genes, the top three in BP, CC and MF were selected for enrichment analysis. Among down-regulated genes, the top three enrichment in BP, CC were selected for analysis, while the top two enrichment in MF were selected for analysis. (D) The up-regulated and down-regulated Genes were analyzed by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. (E) Flow cytometry analysis of ROS in JMJD6 overexpression H460 cells. Representative images are shown on the left and statistical data analysis on the right. (F) The changes of lipid peroxidation level in JMJD6 over-expressed H1299 cells were detected by MDA method. (G) JC-1 fluorescent probe was used to detect the degree of mitochondrial membrane potential depolarization in H1299 cells with JMJD6 overexpression. Representative images are shown on the left and statistical data analysis on the right. The scale bars represent 50 μm. (H) ROS content in JMJD6 knockdown H157 cells was detected by flow cytometry. Representative images are shown on the left and statistical data analysis on the right. (I) MDA assay was used to detect the changes of lipid peroxidation in JMJD6 knockdown A549 cells. (J) JC-1 fluorescent probe was used to detect the changes of mitochondrial membrane potential in JMJD6 knockdown A549 cells. Representative images are shown on the left and statistical data analysis on the right. The scale bars represent 50 μm. (K) The JMJD6 knockdown A549 cells were treated with erastin and fer-1, respectively, and the cell viability was detected. (L, M, N) Tumor images of nude mice in control group, overexpressed JMJD6 group and overexpressed JMJD6 with injected 30 mg/kg erastin group (L). Tumor weight (M) and growth curve (N) were measured. Data are presented as mean ± standard deviation. Data are presented as mean ± standard deviation. *p < 0.05; **p < 0.01; ***p < 0.001

Article Snippet: The antibodies used in this study were listed as follows: JMJD6 antibody (Santa Cruz Biotechnology, Cat# sc-28348, 1:1000), METTL14 antibody (CST, Cat# 48699, 1:1000), SLC3A2 antibody (Abways, Cat# CY8416, 1:1000), GAPDH antibody (Abways, Cat# AB2000, 1:6000), Flag-tag antibody (Abways, Cat# AB0030, 1:1000), HAtag antibody (proteintech, Cat# 51064-2-AP, 1:3000), Acetylated-Lysine Antibody (CST, Cat# 9441, 1:1000), JMJD6-acK375 antibody (Jiaxuan Biotech Cat#JXR7611, 1:1000), Rabbit Anti-Goat IgG (H + L) HRP (Abways, Cat# AB0101, 1:8000).

Techniques: Gene Expression, Expressing, RNA Sequencing, Functional Assay, Flow Cytometry, Over Expression, Membrane, Knockdown, Multiple Displacement Amplification, Control, Injection, Standard Deviation

Journal: eLife

Article Title: Metabolic reprogramming of cancer cells by JMJD6-mediated pre-mRNA splicing associated with therapeutic response to splicing inhibitor

doi: 10.7554/eLife.90993

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-JMJD6 (mouse monoclonal) , Santa Cruz Biotechnology , sc-28348, RRID: AB_628185 , WB 1:1000.

Techniques: Recombinant, SYBR Green Assay, Real-time Polymerase Chain Reaction, Labeling, Transfection, Protease Inhibitor, Immunoprecipitation, RNA Immunoprecipitation, Plasmid Preparation, Expressing, Sequencing, shRNA, Knockdown, CRISPR, Software, Gene Expression, Alternative Splicing

Figure 2. JMJD6 is required for neuroblastoma growth and facilitates MYC-mediated cellular transformation. (A) Copy number of genes encoding JmjC domain proteins in St Jude neuroblastoma cohort (https://platform.stjude.cloud). (B) Kaplan-Meier survival curve showing high JMJD6 is correlated with worse event-free survival (SEQC RNA-seq dataset). (C) Crystal violet showing the colony staining on day 7 after JMJD6 shRNA knockdown in neuroblastoma cell lines validated by western blot (harvested at 72 hr). n=single experiment. (D) Xenograft tumor growth of BE2C (right) models with lentiviral JMJD6 shRNA knockdown. p-Value calculated by multiple unpaired t-test across each row. n=5 per group. ***p<0.001, **p<0.01. (E) Xenograft tumor growth of SK-N-AS models with lentiviral JMJD6 shRNA knockdown. p-value calculated by multiple unpaired t-test across each row. ***p<0.001, **p<0.01. (F) Western blot validating the expression of retroviral-based MYCN and JMJD6 in JoMa1 cells. (G) Cell proliferation of JoMa1 cells transduced with indicated constructs expressing GFP, JMJD6, MYCN, JMJD6+MYCN. (H) Colony formation of JoMa1 cells transduced with indicated constructs, GFP, JMJD6, MYCN, JMJD6+MYCN. Top panel showing photos taken under light microscope. Bottom panel showing cell colonies stained with crystal violet. (I) Xenograft tumor growth of JoMa1 cells transduced with indicated constructs, GFP, JMJD6, MYCN, JMJD6+MYCN. n=5 per group. p-Value calculated by multiple unpaired t-test across each row. ***p<0.001, **p<0.01. Data are Mean ± SEM.

Journal: eLife

Article Title: Metabolic reprogramming of cancer cells by JMJD6-mediated pre-mRNA splicing associated with therapeutic response to splicing inhibitor

doi: 10.7554/elife.90993

Figure Lengend Snippet: Figure 2. JMJD6 is required for neuroblastoma growth and facilitates MYC-mediated cellular transformation. (A) Copy number of genes encoding JmjC domain proteins in St Jude neuroblastoma cohort (https://platform.stjude.cloud). (B) Kaplan-Meier survival curve showing high JMJD6 is correlated with worse event-free survival (SEQC RNA-seq dataset). (C) Crystal violet showing the colony staining on day 7 after JMJD6 shRNA knockdown in neuroblastoma cell lines validated by western blot (harvested at 72 hr). n=single experiment. (D) Xenograft tumor growth of BE2C (right) models with lentiviral JMJD6 shRNA knockdown. p-Value calculated by multiple unpaired t-test across each row. n=5 per group. ***p<0.001, **p<0.01. (E) Xenograft tumor growth of SK-N-AS models with lentiviral JMJD6 shRNA knockdown. p-value calculated by multiple unpaired t-test across each row. ***p<0.001, **p<0.01. (F) Western blot validating the expression of retroviral-based MYCN and JMJD6 in JoMa1 cells. (G) Cell proliferation of JoMa1 cells transduced with indicated constructs expressing GFP, JMJD6, MYCN, JMJD6+MYCN. (H) Colony formation of JoMa1 cells transduced with indicated constructs, GFP, JMJD6, MYCN, JMJD6+MYCN. Top panel showing photos taken under light microscope. Bottom panel showing cell colonies stained with crystal violet. (I) Xenograft tumor growth of JoMa1 cells transduced with indicated constructs, GFP, JMJD6, MYCN, JMJD6+MYCN. n=5 per group. p-Value calculated by multiple unpaired t-test across each row. ***p<0.001, **p<0.01. Data are Mean ± SEM.

Article Snippet: GAPDH (Cell Signaling Technology, 5174s, rabbit antibody), MYCN (Santa Cruz Biotechnology, 53993, mouse antibody), FLAG (Sigma, F1804, mouse antibody), Biotin (Bethyl Laboratories, A150109A, rabbit antibody), ACTIN (Sigma, A3854, mouse antibody), PUF60 (Thermo Fisher, PA5- 21411, rabbit antibody), U2AF2 (Novus Biologicals, NBP2- 04140, rabbit antibody), CPSF6 (Bethyl Laboratories, 357A, rabbit antibody), DHX40 (Novus Biologicals, NBP1- 91834, rabbit antibody), DHX8 (Abcam, AB181074, rabbit antibody), LUC7L1 (Novus Biologicals, NBP2- 56401, rabbit antibody), LUC7L2 (Novus Biologicals, NBP2- 33621, rabbit antibody), LUC7L3 (Novus Biologicals, NBP1- 88053, rabbit antibody), RBM39 (ATLAS, HPA001519, rabbit antibody), GLS (KGA- specific) (Proteintech, 20170- 1- AP, rabbit antibody), GLS (GAC- specific) (Proteintech, 19958- 1- AP, rabbit antibody), JMJD6 (ATLAS, HPA059156, rabbit antibody), JMJD6 (Santa Cruz Biotechnology, sc- 28348, mouse antibody).

Techniques: Transformation Assay, RNA Sequencing, Staining, shRNA, Knockdown, Western Blot, Expressing, Retroviral, Transduction, Construct, Light Microscopy

Figure 3. JMJD6 regulates pre-mRNA splicing of genes involved in metabolism. (A) Pathway enrichment for JMJD6 co-dependency genes whose knockout exhibits similar phenotype with JMJD6 knockout based on re-analysis of DepMap data. (B) Pathway enrichment for genes whose knockout exhibits opposite phenotype with JMJD6 knockout based on re-analysis of DepMap data. (C) Chromosomal location enrichment for JMJD6 co- dependency genes whose knockout exhibits similar phenotype with JMJD6 knockout based on re-analysis of DepMap data. (D) Chromosomal location

Journal: eLife

Article Title: Metabolic reprogramming of cancer cells by JMJD6-mediated pre-mRNA splicing associated with therapeutic response to splicing inhibitor

doi: 10.7554/elife.90993

Figure Lengend Snippet: Figure 3. JMJD6 regulates pre-mRNA splicing of genes involved in metabolism. (A) Pathway enrichment for JMJD6 co-dependency genes whose knockout exhibits similar phenotype with JMJD6 knockout based on re-analysis of DepMap data. (B) Pathway enrichment for genes whose knockout exhibits opposite phenotype with JMJD6 knockout based on re-analysis of DepMap data. (C) Chromosomal location enrichment for JMJD6 co- dependency genes whose knockout exhibits similar phenotype with JMJD6 knockout based on re-analysis of DepMap data. (D) Chromosomal location

Article Snippet: GAPDH (Cell Signaling Technology, 5174s, rabbit antibody), MYCN (Santa Cruz Biotechnology, 53993, mouse antibody), FLAG (Sigma, F1804, mouse antibody), Biotin (Bethyl Laboratories, A150109A, rabbit antibody), ACTIN (Sigma, A3854, mouse antibody), PUF60 (Thermo Fisher, PA5- 21411, rabbit antibody), U2AF2 (Novus Biologicals, NBP2- 04140, rabbit antibody), CPSF6 (Bethyl Laboratories, 357A, rabbit antibody), DHX40 (Novus Biologicals, NBP1- 91834, rabbit antibody), DHX8 (Abcam, AB181074, rabbit antibody), LUC7L1 (Novus Biologicals, NBP2- 56401, rabbit antibody), LUC7L2 (Novus Biologicals, NBP2- 33621, rabbit antibody), LUC7L3 (Novus Biologicals, NBP1- 88053, rabbit antibody), RBM39 (ATLAS, HPA001519, rabbit antibody), GLS (KGA- specific) (Proteintech, 20170- 1- AP, rabbit antibody), GLS (GAC- specific) (Proteintech, 19958- 1- AP, rabbit antibody), JMJD6 (ATLAS, HPA059156, rabbit antibody), JMJD6 (Santa Cruz Biotechnology, sc- 28348, mouse antibody).

Techniques: Knock-Out

Figure 4. JMJD6 regulates alternative splicing of glutaminolysis gene, GLS. (A) Sashimi plot showing the alternative splicing of GLS after JMJD6 knockdown in BE2C cells in duplicates. The number indicates the RNA-seq read counts of exon junction. (B) Real-time (RT)-polymerase chain reaction (PCR) assessing the relative expression of GAC and KGA isoforms after JMJD6 knockdown in BE2C cells in triplicates. (C) Western blot showing the expression of GAC and KGA isoforms in SK-N-AS, BE2C, SIMA after JMJD6 knockdown for 72 hr. (D) KGA- and GAC-specific reporter analysis showing

Journal: eLife

Article Title: Metabolic reprogramming of cancer cells by JMJD6-mediated pre-mRNA splicing associated with therapeutic response to splicing inhibitor

doi: 10.7554/elife.90993

Figure Lengend Snippet: Figure 4. JMJD6 regulates alternative splicing of glutaminolysis gene, GLS. (A) Sashimi plot showing the alternative splicing of GLS after JMJD6 knockdown in BE2C cells in duplicates. The number indicates the RNA-seq read counts of exon junction. (B) Real-time (RT)-polymerase chain reaction (PCR) assessing the relative expression of GAC and KGA isoforms after JMJD6 knockdown in BE2C cells in triplicates. (C) Western blot showing the expression of GAC and KGA isoforms in SK-N-AS, BE2C, SIMA after JMJD6 knockdown for 72 hr. (D) KGA- and GAC-specific reporter analysis showing

Article Snippet: GAPDH (Cell Signaling Technology, 5174s, rabbit antibody), MYCN (Santa Cruz Biotechnology, 53993, mouse antibody), FLAG (Sigma, F1804, mouse antibody), Biotin (Bethyl Laboratories, A150109A, rabbit antibody), ACTIN (Sigma, A3854, mouse antibody), PUF60 (Thermo Fisher, PA5- 21411, rabbit antibody), U2AF2 (Novus Biologicals, NBP2- 04140, rabbit antibody), CPSF6 (Bethyl Laboratories, 357A, rabbit antibody), DHX40 (Novus Biologicals, NBP1- 91834, rabbit antibody), DHX8 (Abcam, AB181074, rabbit antibody), LUC7L1 (Novus Biologicals, NBP2- 56401, rabbit antibody), LUC7L2 (Novus Biologicals, NBP2- 33621, rabbit antibody), LUC7L3 (Novus Biologicals, NBP1- 88053, rabbit antibody), RBM39 (ATLAS, HPA001519, rabbit antibody), GLS (KGA- specific) (Proteintech, 20170- 1- AP, rabbit antibody), GLS (GAC- specific) (Proteintech, 19958- 1- AP, rabbit antibody), JMJD6 (ATLAS, HPA059156, rabbit antibody), JMJD6 (Santa Cruz Biotechnology, sc- 28348, mouse antibody).

Techniques: Alternative Splicing, Knockdown, RNA Sequencing, Polymerase Chain Reaction, Expressing, Western Blot

Figure 6. JMJD6 forms an interaction network that consists of proteins involved in splicing and protein synthesis. (A) FLAG-tagged JMJD6 transduced into SK-N-AS cells for immunoprecipitation with anti-FLAG followed by protein identification by mass spectrometry. The interacting protein partners of JMJD6 are analyzed by STRING protein network. (B) Immunoprecipitation followed by western blot to validate the JMJD6-interacting partners in SK-N-AS cells. IP = immunoprecipitation, FT = flowthrough. n=single experiment. (C) Click-iT AHA labeling showing the newly synthesized proteins after overexpression of JMJD6 in SK-N-AS cells. n=single experiment. (D, E) Western blot showing the expression of GAC and KGA isoforms in SKNAS (D), BE2C (E), after U2AF2 and CPSF6 knockdown for 72 hr. n=single experiment.

Journal: eLife

Article Title: Metabolic reprogramming of cancer cells by JMJD6-mediated pre-mRNA splicing associated with therapeutic response to splicing inhibitor

doi: 10.7554/elife.90993

Figure Lengend Snippet: Figure 6. JMJD6 forms an interaction network that consists of proteins involved in splicing and protein synthesis. (A) FLAG-tagged JMJD6 transduced into SK-N-AS cells for immunoprecipitation with anti-FLAG followed by protein identification by mass spectrometry. The interacting protein partners of JMJD6 are analyzed by STRING protein network. (B) Immunoprecipitation followed by western blot to validate the JMJD6-interacting partners in SK-N-AS cells. IP = immunoprecipitation, FT = flowthrough. n=single experiment. (C) Click-iT AHA labeling showing the newly synthesized proteins after overexpression of JMJD6 in SK-N-AS cells. n=single experiment. (D, E) Western blot showing the expression of GAC and KGA isoforms in SKNAS (D), BE2C (E), after U2AF2 and CPSF6 knockdown for 72 hr. n=single experiment.

Article Snippet: GAPDH (Cell Signaling Technology, 5174s, rabbit antibody), MYCN (Santa Cruz Biotechnology, 53993, mouse antibody), FLAG (Sigma, F1804, mouse antibody), Biotin (Bethyl Laboratories, A150109A, rabbit antibody), ACTIN (Sigma, A3854, mouse antibody), PUF60 (Thermo Fisher, PA5- 21411, rabbit antibody), U2AF2 (Novus Biologicals, NBP2- 04140, rabbit antibody), CPSF6 (Bethyl Laboratories, 357A, rabbit antibody), DHX40 (Novus Biologicals, NBP1- 91834, rabbit antibody), DHX8 (Abcam, AB181074, rabbit antibody), LUC7L1 (Novus Biologicals, NBP2- 56401, rabbit antibody), LUC7L2 (Novus Biologicals, NBP2- 33621, rabbit antibody), LUC7L3 (Novus Biologicals, NBP1- 88053, rabbit antibody), RBM39 (ATLAS, HPA001519, rabbit antibody), GLS (KGA- specific) (Proteintech, 20170- 1- AP, rabbit antibody), GLS (GAC- specific) (Proteintech, 19958- 1- AP, rabbit antibody), JMJD6 (ATLAS, HPA059156, rabbit antibody), JMJD6 (Santa Cruz Biotechnology, sc- 28348, mouse antibody).

Techniques: Immunoprecipitation, Mass Spectrometry, Western Blot, Labeling, Synthesized, Over Expression, Expressing, Knockdown

Figure 7. JMJD6 regulates production of citric acid cycle intermediates and NTP. (A) Heatmap showing the metabolites differentially expressed in SK- N-AS cells (n=5) after JMJD6 knockout (n=5) based on liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis. (B) Pathway analysis of metabolites downregulated by JMJD6 knockout. (C) Pathway cartoon showing the connections of tricarboxylic acid (TCA), glycolysis, glutaminolysis, and β-oxidation. (D) Correlation of metabolite abundance with JMJD6 dependency. The positive correlation indicates that the higher the abundance of metabolites, the more resistance of cells to JMJD6 knockout. On the contrary, the negative correlation indicates the higher the abundance of metabolites, the more sensitive of cells to JMJD6 knockout.

Journal: eLife

Article Title: Metabolic reprogramming of cancer cells by JMJD6-mediated pre-mRNA splicing associated with therapeutic response to splicing inhibitor

doi: 10.7554/elife.90993

Figure Lengend Snippet: Figure 7. JMJD6 regulates production of citric acid cycle intermediates and NTP. (A) Heatmap showing the metabolites differentially expressed in SK- N-AS cells (n=5) after JMJD6 knockout (n=5) based on liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis. (B) Pathway analysis of metabolites downregulated by JMJD6 knockout. (C) Pathway cartoon showing the connections of tricarboxylic acid (TCA), glycolysis, glutaminolysis, and β-oxidation. (D) Correlation of metabolite abundance with JMJD6 dependency. The positive correlation indicates that the higher the abundance of metabolites, the more resistance of cells to JMJD6 knockout. On the contrary, the negative correlation indicates the higher the abundance of metabolites, the more sensitive of cells to JMJD6 knockout.

Article Snippet: GAPDH (Cell Signaling Technology, 5174s, rabbit antibody), MYCN (Santa Cruz Biotechnology, 53993, mouse antibody), FLAG (Sigma, F1804, mouse antibody), Biotin (Bethyl Laboratories, A150109A, rabbit antibody), ACTIN (Sigma, A3854, mouse antibody), PUF60 (Thermo Fisher, PA5- 21411, rabbit antibody), U2AF2 (Novus Biologicals, NBP2- 04140, rabbit antibody), CPSF6 (Bethyl Laboratories, 357A, rabbit antibody), DHX40 (Novus Biologicals, NBP1- 91834, rabbit antibody), DHX8 (Abcam, AB181074, rabbit antibody), LUC7L1 (Novus Biologicals, NBP2- 56401, rabbit antibody), LUC7L2 (Novus Biologicals, NBP2- 33621, rabbit antibody), LUC7L3 (Novus Biologicals, NBP1- 88053, rabbit antibody), RBM39 (ATLAS, HPA001519, rabbit antibody), GLS (KGA- specific) (Proteintech, 20170- 1- AP, rabbit antibody), GLS (GAC- specific) (Proteintech, 19958- 1- AP, rabbit antibody), JMJD6 (ATLAS, HPA059156, rabbit antibody), JMJD6 (Santa Cruz Biotechnology, sc- 28348, mouse antibody).

Techniques: Knock-Out, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy

Figure 8. JMJD6-GAC pathway regulates the response of neuroblastoma cells to indisulam treatment. (A) Spearman correlation of effects of JMJD6 knockout and RBM39 knockout demonstrating the co-dependency of JMJD6 and RBM39 from DepMap CRISPR screening data (n=1086). Each dot represents one cell line. (B) Gene set enrichment analysis (GSEA) for indisulam sensitive vs resistant neuroblastoma cell lines based on CTD2 (Cancer Target Discovery and Development) data showing histone lysine demethylase gene signature is the one that is significantly associated with indisulam response. (C) Heatmap from GSEA (B) showing the individual genes in indisulam-sensitive and -resistant cells. (D) JMJD6 expression in indisulam- sensitive and -resistant neuroblastoma cells. p-Value calculated by Student’s t-test. (E) Western blot showing JMJD6 knockout in SK-N-AS cells using indicated antibodies. (F) Colony formation of SK-N-AS cells in triplicates with or without JMJD6 knockout treated with different concentrations of indisulam for 7 days, stained with crystal violet. n=3 per group. (G) Quantification of cell density by using ImageJ software from (F) (n=triplicates). ns = not significant. **p<0.001, ***p<0.0001. (H) Western blot showing JMJD6 knockout in BE2C cells using indicated antibodies. (I) Colony formation of BE2C cells in triplicates with or without JMJD6 knockout treated with 100 nM of indisulam for 5 days, stained with crystal violet. n=3 per group. (J) Quantification of cell density by using ImageJ software from (I) (n=triplicates). ns = not significant. **p<0.001, ***p<0.0001. (K) Colony formation

Journal: eLife

Article Title: Metabolic reprogramming of cancer cells by JMJD6-mediated pre-mRNA splicing associated with therapeutic response to splicing inhibitor

doi: 10.7554/elife.90993

Figure Lengend Snippet: Figure 8. JMJD6-GAC pathway regulates the response of neuroblastoma cells to indisulam treatment. (A) Spearman correlation of effects of JMJD6 knockout and RBM39 knockout demonstrating the co-dependency of JMJD6 and RBM39 from DepMap CRISPR screening data (n=1086). Each dot represents one cell line. (B) Gene set enrichment analysis (GSEA) for indisulam sensitive vs resistant neuroblastoma cell lines based on CTD2 (Cancer Target Discovery and Development) data showing histone lysine demethylase gene signature is the one that is significantly associated with indisulam response. (C) Heatmap from GSEA (B) showing the individual genes in indisulam-sensitive and -resistant cells. (D) JMJD6 expression in indisulam- sensitive and -resistant neuroblastoma cells. p-Value calculated by Student’s t-test. (E) Western blot showing JMJD6 knockout in SK-N-AS cells using indicated antibodies. (F) Colony formation of SK-N-AS cells in triplicates with or without JMJD6 knockout treated with different concentrations of indisulam for 7 days, stained with crystal violet. n=3 per group. (G) Quantification of cell density by using ImageJ software from (F) (n=triplicates). ns = not significant. **p<0.001, ***p<0.0001. (H) Western blot showing JMJD6 knockout in BE2C cells using indicated antibodies. (I) Colony formation of BE2C cells in triplicates with or without JMJD6 knockout treated with 100 nM of indisulam for 5 days, stained with crystal violet. n=3 per group. (J) Quantification of cell density by using ImageJ software from (I) (n=triplicates). ns = not significant. **p<0.001, ***p<0.0001. (K) Colony formation

Article Snippet: GAPDH (Cell Signaling Technology, 5174s, rabbit antibody), MYCN (Santa Cruz Biotechnology, 53993, mouse antibody), FLAG (Sigma, F1804, mouse antibody), Biotin (Bethyl Laboratories, A150109A, rabbit antibody), ACTIN (Sigma, A3854, mouse antibody), PUF60 (Thermo Fisher, PA5- 21411, rabbit antibody), U2AF2 (Novus Biologicals, NBP2- 04140, rabbit antibody), CPSF6 (Bethyl Laboratories, 357A, rabbit antibody), DHX40 (Novus Biologicals, NBP1- 91834, rabbit antibody), DHX8 (Abcam, AB181074, rabbit antibody), LUC7L1 (Novus Biologicals, NBP2- 56401, rabbit antibody), LUC7L2 (Novus Biologicals, NBP2- 33621, rabbit antibody), LUC7L3 (Novus Biologicals, NBP1- 88053, rabbit antibody), RBM39 (ATLAS, HPA001519, rabbit antibody), GLS (KGA- specific) (Proteintech, 20170- 1- AP, rabbit antibody), GLS (GAC- specific) (Proteintech, 19958- 1- AP, rabbit antibody), JMJD6 (ATLAS, HPA059156, rabbit antibody), JMJD6 (Santa Cruz Biotechnology, sc- 28348, mouse antibody).

Techniques: Knock-Out, CRISPR, Expressing, Western Blot, Staining, Software

Figure 9. Working mechanism of JMJD6 in MYC-driven neuroblastoma. Overactive MYC drives high-load of gene transcription, enhanced protein synthesis, and high rate of metabolism, leading to detrimental cellular stresses and consequent cell death (Model A). However, when 17q is amplified, high levels of JMJD6 and other proteins encoded by 17q genes physically interact with the splicing and translational machineries, enhancing pre-mRNA splicing of metabolic genes such as glutaminase (GLS) and inhibiting global protein synthesis, respectively, leading to reduced detrimental stresses and enhanced cancer cell survival and tumorigenesis (Model B). The high levels of JMJD6 predicts high dependency of RBM39, which are more sensitive to indisulam treatment.

Journal: eLife

Article Title: Metabolic reprogramming of cancer cells by JMJD6-mediated pre-mRNA splicing associated with therapeutic response to splicing inhibitor

doi: 10.7554/elife.90993

Figure Lengend Snippet: Figure 9. Working mechanism of JMJD6 in MYC-driven neuroblastoma. Overactive MYC drives high-load of gene transcription, enhanced protein synthesis, and high rate of metabolism, leading to detrimental cellular stresses and consequent cell death (Model A). However, when 17q is amplified, high levels of JMJD6 and other proteins encoded by 17q genes physically interact with the splicing and translational machineries, enhancing pre-mRNA splicing of metabolic genes such as glutaminase (GLS) and inhibiting global protein synthesis, respectively, leading to reduced detrimental stresses and enhanced cancer cell survival and tumorigenesis (Model B). The high levels of JMJD6 predicts high dependency of RBM39, which are more sensitive to indisulam treatment.

Article Snippet: GAPDH (Cell Signaling Technology, 5174s, rabbit antibody), MYCN (Santa Cruz Biotechnology, 53993, mouse antibody), FLAG (Sigma, F1804, mouse antibody), Biotin (Bethyl Laboratories, A150109A, rabbit antibody), ACTIN (Sigma, A3854, mouse antibody), PUF60 (Thermo Fisher, PA5- 21411, rabbit antibody), U2AF2 (Novus Biologicals, NBP2- 04140, rabbit antibody), CPSF6 (Bethyl Laboratories, 357A, rabbit antibody), DHX40 (Novus Biologicals, NBP1- 91834, rabbit antibody), DHX8 (Abcam, AB181074, rabbit antibody), LUC7L1 (Novus Biologicals, NBP2- 56401, rabbit antibody), LUC7L2 (Novus Biologicals, NBP2- 33621, rabbit antibody), LUC7L3 (Novus Biologicals, NBP1- 88053, rabbit antibody), RBM39 (ATLAS, HPA001519, rabbit antibody), GLS (KGA- specific) (Proteintech, 20170- 1- AP, rabbit antibody), GLS (GAC- specific) (Proteintech, 19958- 1- AP, rabbit antibody), JMJD6 (ATLAS, HPA059156, rabbit antibody), JMJD6 (Santa Cruz Biotechnology, sc- 28348, mouse antibody).

Techniques: Amplification

Fig. 4 Tumor-CM activates THP-1 cells and upregulates JMJD6 expression. a Morphology of THP-1 cells cultured in normal medium, with or without PMA and tumor-CM (H460) for 48 h. Scale bar, 100 μm. b The changes of macrophage related markers on the surface of THP-1 cells were detected by flow cytometry after PMA induction. Representative results. c CD206 expression of TAM surfaces were detected by flow cytometry after tumor-CM induction. d The secretion of IL-10 in the TAM culture supernatant detected by CBA assays. e THP-1 cells were treated with PMA alone or tumor-CM for 2 or 3 days, followed by JMJD6 protein level detection. f Representative images of immunofluorescence staining of CD68 (red), JMJD6 (green) and DAPI (blue) in THP-1 cells treated with PMA alone or tumor-CM for 2 days, the scale bars 10 μm and 20 μm (right panel) as indicated in figures. Data are shown as mean ± SD (n = 3). *P < 0.05, **P < 0.01.

Journal: Oncogene

Article Title: JMJD6 in tumor-associated macrophage regulates macrophage polarization and cancer progression via STAT3/IL-10 axis.

doi: 10.1038/s41388-023-02781-9

Figure Lengend Snippet: Fig. 4 Tumor-CM activates THP-1 cells and upregulates JMJD6 expression. a Morphology of THP-1 cells cultured in normal medium, with or without PMA and tumor-CM (H460) for 48 h. Scale bar, 100 μm. b The changes of macrophage related markers on the surface of THP-1 cells were detected by flow cytometry after PMA induction. Representative results. c CD206 expression of TAM surfaces were detected by flow cytometry after tumor-CM induction. d The secretion of IL-10 in the TAM culture supernatant detected by CBA assays. e THP-1 cells were treated with PMA alone or tumor-CM for 2 or 3 days, followed by JMJD6 protein level detection. f Representative images of immunofluorescence staining of CD68 (red), JMJD6 (green) and DAPI (blue) in THP-1 cells treated with PMA alone or tumor-CM for 2 days, the scale bars 10 μm and 20 μm (right panel) as indicated in figures. Data are shown as mean ± SD (n = 3). *P < 0.05, **P < 0.01.

Article Snippet: The antibodies we used: Anti-JMJD6: sc-28348 (human), sc-28349 (mouse); Anti-GAPDH: sc-32233, Santa Cruz, CA, USA; Anti-STAT3: 9139, CST; Anti-p-STAT3Y705: 145, CST; Anti-F4/80: ab16911, abcam; Anti-IL-10:ER1911-19; Anti-CD31: GB11063; Anti-Ki67: GB111499; Goat anti-Rabbit IgG (H+ L) Secondary Antibody, HRP: Invitrogen, 31460; Goat anti-Mouse IgG (H+ L) Secondary Antibody, HRP: Invitrogen, 31430.

Techniques: Expressing, Cell Culture, Cytometry, Staining

Fig. 7 JMJD6 knockdown enhances PD-1-sensitivity in mice. a Schematic diagram of anti-PD-1 treatment on WT and Jmjd6+/−mice B16F10 lung metastasis tumor model. b Lung photo at the end of the experiment. c Statistical results of the number of lung tumor nodules in mice. d Representative images of mouse lungs H&E staining. e The infiltration of M2-like TAMs (CD206+ MHC-II-), detected by flow cytometry. f The infiltration of CD3+ total T cells, detected by flow cytometry. g The infiltration of CD8+ GZMB+ T cells, detected by flow cytometry. h Illustration of mechanism for JMJD6/STAT3/IL-10 axis. Data represent mean ± SD (n = 5). ns: no significant difference, *P < 0.05, **P < 0.01, ***P < 0.001.

Journal: Oncogene

Article Title: JMJD6 in tumor-associated macrophage regulates macrophage polarization and cancer progression via STAT3/IL-10 axis.

doi: 10.1038/s41388-023-02781-9

Figure Lengend Snippet: Fig. 7 JMJD6 knockdown enhances PD-1-sensitivity in mice. a Schematic diagram of anti-PD-1 treatment on WT and Jmjd6+/−mice B16F10 lung metastasis tumor model. b Lung photo at the end of the experiment. c Statistical results of the number of lung tumor nodules in mice. d Representative images of mouse lungs H&E staining. e The infiltration of M2-like TAMs (CD206+ MHC-II-), detected by flow cytometry. f The infiltration of CD3+ total T cells, detected by flow cytometry. g The infiltration of CD8+ GZMB+ T cells, detected by flow cytometry. h Illustration of mechanism for JMJD6/STAT3/IL-10 axis. Data represent mean ± SD (n = 5). ns: no significant difference, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The antibodies we used: Anti-JMJD6: sc-28348 (human), sc-28349 (mouse); Anti-GAPDH: sc-32233, Santa Cruz, CA, USA; Anti-STAT3: 9139, CST; Anti-p-STAT3Y705: 145, CST; Anti-F4/80: ab16911, abcam; Anti-IL-10:ER1911-19; Anti-CD31: GB11063; Anti-Ki67: GB111499; Goat anti-Rabbit IgG (H+ L) Secondary Antibody, HRP: Invitrogen, 31460; Goat anti-Mouse IgG (H+ L) Secondary Antibody, HRP: Invitrogen, 31430.

Techniques: Knockdown, Staining, Cytometry