Journal: Cellular immunology

Article Title: Zinc finger protein 217 contributes to natural killer cell dysfunction in murine colorectal cancer.

doi: 10.1016/j.cellimm.2025.104971

Figure Lengend Snippet: Fig. 3. ZNF217 expression in NK cells. (A) ZNF217 protein quantification in NK cells. Upper panel: a representative immunoblotting image. Lower panel: relative ZNF217 protein expression. w0: week 0. w8: week 8 after AOM injection. w16: week 16 after AOM injection. (B) ZNF217 mRNA levels in LAG-3−and LAG-3+ NK cells. (C) ZNF217 mRNA levels in TIGIT−and TIGIT+ NK cells. *: P < 0.05. **: P < 0.01. ***: P < 0.001. N = 4 or 6 independent samples. In (A), each sample contains NK cells pooled from fifteen to twenty mice. In (B and C), each sample represents an individual mouse. One-way ANOVA.

Article Snippet: After the standard transfer and blocking procedures, the membrane was incubated with the polyclonal rabbit anti-ZNF217 antibody (Cat# NBP2–20978, Novus Biologicals, 1:1000) or monoclonal rabbit anti-GAPDH antibody (Cat# ab181602, Abcam, 1:5000) at 4 ◦C overnight.

Techniques: Expressing, Western Blot, Injection

Journal: Cellular immunology

Article Title: Zinc finger protein 217 contributes to natural killer cell dysfunction in murine colorectal cancer.

doi: 10.1016/j.cellimm.2025.104971

Figure Lengend Snippet: Fig. 4. The effect of ZNF217 knockdown on NK cell activity. (A) Representative density plots illustrating GFP expression on day 2 after lentivirus transduction. Un: Untreated NK cells. SCR: Scramble control virus transduction. SiZ: ZNF217 shRNA virus transduction. (B) ZNF217 mRNA levels in NK cells on day 2 after viral transduction. Un: Untreated NK cells. cNK: control GFP+ NK cells. zNK: GFP+ ZNF217-knockdown NK cells. (C) Representative density plots illustrating NK cell apoptosis and necrosis after transduction. (D) The proportions of dead (apoptotic plus necrotic) NK cells. (E) Representative histograms illustrating Ki67 expression in NK cells. (F) The proportions of Ki67+ NK cells. (G) Representative density plots illustrating the expression of granzyme B (GZMB) and perforin (PFR) in NK cells. *: P < 0.05. **: P < 0.01. ns: not statistically significant. N = 6 independent samples. One-way ANOVA.

Article Snippet: After the standard transfer and blocking procedures, the membrane was incubated with the polyclonal rabbit anti-ZNF217 antibody (Cat# NBP2–20978, Novus Biologicals, 1:1000) or monoclonal rabbit anti-GAPDH antibody (Cat# ab181602, Abcam, 1:5000) at 4 ◦C overnight.

Techniques: Knockdown, Activity Assay, Expressing, Transduction, Control, Virus, shRNA

Journal: Cellular immunology

Article Title: Zinc finger protein 217 contributes to natural killer cell dysfunction in murine colorectal cancer.

doi: 10.1016/j.cellimm.2025.104971

Figure Lengend Snippet: Fig. 5. The effect of ZNF217 knockdown on NK cell-mediated cytotoxicity. (A) Representative density plots illustrating the expression of LAMP-1 and IFN-γ in NK cells after 4-h co-culture with YAC-1 cells. cNK: control NK cells. zNK: ZNF217-knockdown NK cells. Alone: NK cells alone. With YAC-1: NK cell-YAC-1 cell co-culture. (B and C) The proportions of LAMP-1+ (B) and IFN-γ+ (C) NK cells. (D) Representative density plots illustrating YAC-1 cell apoptosis and necrosis after 4-h co-culture with NK cells. Alone: YAC-1 cell alone. +cNK: YAC-1 cells with control NK cells. +zNK: YAC-1 cells with ZNF217-knockdown NK cells. (E) The proportions of dead (apoptotic plus necrotic) YAC-1 cells. *: P < 0.05. **: P < 0.01. ***: P < 0.001. ns: not statistically significant. N = 6 independent samples. One-way ANOVA.

Article Snippet: After the standard transfer and blocking procedures, the membrane was incubated with the polyclonal rabbit anti-ZNF217 antibody (Cat# NBP2–20978, Novus Biologicals, 1:1000) or monoclonal rabbit anti-GAPDH antibody (Cat# ab181602, Abcam, 1:5000) at 4 ◦C overnight.

Techniques: Knockdown, Expressing, Co-Culture Assay, Control

Journal: Cellular immunology

Article Title: Zinc finger protein 217 contributes to natural killer cell dysfunction in murine colorectal cancer.

doi: 10.1016/j.cellimm.2025.104971

Figure Lengend Snippet: Fig. 6. The effect of ZNF217 knockdown on NK function in vivo. (A) Exogenous GFP+ NK cells in CRC-associated mononuclear cells. Left panel: representative dot plots. Right panel: statistics of exogenous NK cell frequencies. cNK: control NK cells. zNK: ZNF217-knockdown NK cells. (B) TIGIT and Tim-3 expression on exogenous NK cells. Left panel: representative density plots. Right panel: statistics of the frequencies of TIGIT+ and Tim-3+ NK cells. (C) Granzyme B and perforin expression in exogenous NK cells. Left panel: representative density plots. Right panel: statistics of the frequencies of granzyme B+ and perforin+ NK cells. (D) LAMP- 1 and IFN-γ expression in exogenous NK cells. Left panel: representative density plots. Right panel: statistics of the frequencies of LAMP-1+ and IFN-γ+ NK cells. (E) Representative dot plots showing the sequential gating strategy for analyzing the apoptosis and necrosis of primary CRC cells. Saline: Saline infusion. cNK: control NK cell transfer. zNK: ZNF217-knockdown NK cell transfer. (F) Statistics for (E). (G) Tumor size. (H) Histological score. **: P < 0.01. ***: P < 0.001. N = 8 mice. Student's t-test for (A to D). One-way ANOVA for (F to H).

Article Snippet: After the standard transfer and blocking procedures, the membrane was incubated with the polyclonal rabbit anti-ZNF217 antibody (Cat# NBP2–20978, Novus Biologicals, 1:1000) or monoclonal rabbit anti-GAPDH antibody (Cat# ab181602, Abcam, 1:5000) at 4 ◦C overnight.

Techniques: Knockdown, In Vivo, Control, Expressing, Saline

Journal: Cellular immunology

Article Title: Zinc finger protein 217 contributes to natural killer cell dysfunction in murine colorectal cancer.

doi: 10.1016/j.cellimm.2025.104971

Figure Lengend Snippet: Fig. 7. The effect of exhaustion or hypoxia on ZNF217 expression. (A) Representative density plots illustrating the expression of Tim-3 and TIGIT on NK cells after exhaustion induction. Isotype: continuous exposure to isotype antibody. Exhausted: continuous exposure to activating agents. (B and C) Proportions of Tim-3+ (B) and TIGIT+ (C) NK cells. (D) Representative density plots illustrating LAMP-1 and IFN-γ expression in NK cells after exhaustion induction followed by 4-h co-culture with YAC-1 cells. (E and F) Proportions of LAMP-1+ (E) and IFN-γ+ (F) NK cells. (G) ZNF217 mRNA levels in non-exhausted and exhausted NK cells, respectively. (H) ZNF217 mRNA levels in NK cells after 24- and 48-h hypoxia. (I) HIF-1α mRNA levels in NK cells after hypoxia. (J) ZNF217 mRNA levels in NK cells after 24-h normoxia or hypoxia with or without Digoxin. Hypoxia V: addition of vehicle (DMSO) before hypoxia. Hypoxia D: addition of Digoxin before hypoxia. (K) Den sity plots illustrating Granzyme B and perforin expression in NK cells after 48-h hypoxia. cNK: control NK cells. zNK: ZNF217-knockdown NK cells. (L) Proportions of Granzyme B+ (Left panel) and perforin+ (Right panel) NK cells after 48-h hypoxia. (M) Density plots illustrating LAMP-1 and IFN-γ expression in normoxia- or hypoxia-experienced NK cells after culture alone or 4-h co-culture with YAC-1 cells. (N) Proportions of LAMP-1+ (Upper panel) and IFN-γ+ (Lower panel) NK cells for (M). *: P < 0.05. **: P < 0.01. ***: P < 0.001. ns: not statistically significant. N = 3 or 6 independent samples. Student's t-test for (B, C, E, F, G) and One-way ANOVA for (H, I, J, L, and M).

Article Snippet: After the standard transfer and blocking procedures, the membrane was incubated with the polyclonal rabbit anti-ZNF217 antibody (Cat# NBP2–20978, Novus Biologicals, 1:1000) or monoclonal rabbit anti-GAPDH antibody (Cat# ab181602, Abcam, 1:5000) at 4 ◦C overnight.

Techniques: Expressing, Co-Culture Assay, Control, Knockdown

Journal: International Journal of Biological Sciences

Article Title: Core transcriptional regulatory circuitry molecule ZNF217 promotes AML cell proliferation by up-regulating MYB

doi: 10.7150/ijbs.103211

Figure Lengend Snippet: ZNF217, a super-enhancer-regulated gene, exhibits high expression in AML and is correlated with an unfavorable prognosis. A. A heatmap was generated to visualize the occurrence (1 or 0) of SEs in AML cell strains and patient samples. The analysis included samples with a frequency exceeding 80% and fragment lengths greater than 2500bp. The H3K27ac activity profile of SE-driven oncogenic genes on chr20:53592377-53594956, which ranked first, was displayed in 11 AML patient samples (blue tracks) and 7 AML cell lines (red tracks) on the right side. SE regions are indicated by underlines. B. The ZNF217 expression is significantly upregulated in AML. C. Kaplan-Meier curves were employed to analyze the overall survival probability of AML patients based on ZNF217 expression. D. RNA-Seq reveals a significant elevation in ZNF217 mRNA expression within bone marrow cells from pediatric AML patients compared to that in healthy children's bone marrow HSPCs. E. qRT-PCR reveals a significant elevation in ZNF217 mRNA expression within bone marrow cells from pediatric AML patients compared to that in healthy children's bone marrow HSPCs. F. Interactions between the enhancers and promoter region of ZNF217 were observed in MV4-11 and THP1 cells.

Article Snippet: Next, we followed the flow cytometry intracellular staining protocol ( https://assets.thermofisher.com/TFS-Assets/BID/Technical-Notes/flow-intracellular-staining-en.pdf ) and used ZNF217 antibody along with FITC-conjugated secondary antibody (ThermoFisher, A-11034) to simultaneously detect ZNF217 protein expression levels in HSPCs.

Techniques: Expressing, Generated, Activity Assay, RNA Sequencing, Quantitative RT-PCR

Journal: International Journal of Biological Sciences

Article Title: Core transcriptional regulatory circuitry molecule ZNF217 promotes AML cell proliferation by up-regulating MYB

doi: 10.7150/ijbs.103211

Figure Lengend Snippet: ZNF217, ELF1, MEF2D, RUNX2, and FOXP1 co-occupy the SE region of ZNF217 to regulate its expression. A. A WashU plot of CUT&Tag data illustrates the co-occupancy (shaded) of ZNF217, ELF1, MEF2D, RUNX2, and FOXP1 at the SEs regions of ZNF217. B. A heatmap was generated to depict the Pearson correlation coefficient between candidate TFs in AML (n=173). C. A heatmap was created to display the fold changes in mRNA levels of candidate TFs after knockdown of each TF using shRNA in MV4-11 cell line. D. Western blotting analysis was performed to validate the coregulation among candidate TFs in MV4-11 cells. E. The luciferase activity of E1 were measured following shRNA knockdown of each candidate TF in MV4-11 cells. F. A schematic diagram depicts the mode of co-regulation of ZNF217 by the five candidate TFs through enhancer1 (E1).

Article Snippet: Next, we followed the flow cytometry intracellular staining protocol ( https://assets.thermofisher.com/TFS-Assets/BID/Technical-Notes/flow-intracellular-staining-en.pdf ) and used ZNF217 antibody along with FITC-conjugated secondary antibody (ThermoFisher, A-11034) to simultaneously detect ZNF217 protein expression levels in HSPCs.

Techniques: Expressing, Generated, Knockdown, shRNA, Western Blot, Luciferase, Activity Assay

Journal: International Journal of Biological Sciences

Article Title: Core transcriptional regulatory circuitry molecule ZNF217 promotes AML cell proliferation by up-regulating MYB

doi: 10.7150/ijbs.103211

Figure Lengend Snippet: Knockdown of ZNF217 suppresses AML proliferation in vitro . A. The expression of ZNF217 was assessed in AML cell lines using Western blotting. B. The efficiency of ZNF217 knockdown in AML cells was evaluated by qRT-PCR. C. The expression levels of ZNF217, c-Myc, CDK2, PARP, and GAPDH in AML cells were examined by Western blotting analysis. D-E. Colony formation test was carried out to evaluate the impact of ZNF217 on colony formation. F. The CCK8 assay was used to determine the proliferation of AML cells transfected with sh-NC or sh-ZNF217. G. Knockdown of ZNF217 led to block cell cycle at the G1 phase in AML cells. H. Knockdown of ZNF217 led to an increase in apoptotic rates in AML cells.

Article Snippet: Next, we followed the flow cytometry intracellular staining protocol ( https://assets.thermofisher.com/TFS-Assets/BID/Technical-Notes/flow-intracellular-staining-en.pdf ) and used ZNF217 antibody along with FITC-conjugated secondary antibody (ThermoFisher, A-11034) to simultaneously detect ZNF217 protein expression levels in HSPCs.

Techniques: Knockdown, In Vitro, Expressing, Western Blot, Quantitative RT-PCR, CCK-8 Assay, Transfection, Blocking Assay

Journal: International Journal of Biological Sciences

Article Title: Core transcriptional regulatory circuitry molecule ZNF217 promotes AML cell proliferation by up-regulating MYB

doi: 10.7150/ijbs.103211

Figure Lengend Snippet: Knockdown of ZNF217 suppresses AML proliferation in vivo . A. Schematic diagram illustrating the animal experimental model. B-C. Representative bioluminescence imaging and statistical analysis at various time points comparing ZNF217 knockdown group with the control group. D. The survival curves were presented in the two groups of mice. E-F. Bioluminescence imaging and statistical analysis of liver/spleen in ZNF217 knockdown group in comparison to the control group. G. Comparison of the percentage of CD45 + cells in bone marrow/liver/spleen tissues between ZNF217 knockdown group and the control group. H. HE staining assessment of bone marrow, liver, and spleen in the two groups of mice. I. Representative images of IHC staining (Ki67) in mice bone marrow.

Article Snippet: Next, we followed the flow cytometry intracellular staining protocol ( https://assets.thermofisher.com/TFS-Assets/BID/Technical-Notes/flow-intracellular-staining-en.pdf ) and used ZNF217 antibody along with FITC-conjugated secondary antibody (ThermoFisher, A-11034) to simultaneously detect ZNF217 protein expression levels in HSPCs.

Techniques: Knockdown, In Vivo, Imaging, Control, Comparison, Staining, Immunohistochemistry

Journal: International Journal of Biological Sciences

Article Title: Core transcriptional regulatory circuitry molecule ZNF217 promotes AML cell proliferation by up-regulating MYB

doi: 10.7150/ijbs.103211

Figure Lengend Snippet: Changes in gene expression following ZNF217 knockout. A-B. RNA-Seq analysis, depicted via volcano plot, revealed the differential expression of genes in MV4-11 and U937 cells between the ZNF217 knockout group and the control group. C-D. GSEA analysis of Hallmark pathways revealed significant enrichment of differentially expressed genes in both cell lines, particularly within signaling pathways associated with MYC, showing negative NES scores indicative of downregulation. E. The heatmap illustrates the top differentially expressed genes in MV4-11 and U937 cells subjected to ZNF217 knockout. F. The bar graph depicts changes in mRNA levels of ZNF217, MYB, HSPD1, CHI3L1, ZNF117, and CD36 subsequent to ZNF217 silencing in MV4-11 and U937 cell lines.

Article Snippet: Next, we followed the flow cytometry intracellular staining protocol ( https://assets.thermofisher.com/TFS-Assets/BID/Technical-Notes/flow-intracellular-staining-en.pdf ) and used ZNF217 antibody along with FITC-conjugated secondary antibody (ThermoFisher, A-11034) to simultaneously detect ZNF217 protein expression levels in HSPCs.

Techniques: Gene Expression, Knock-Out, RNA Sequencing, Quantitative Proteomics, Control, Protein-Protein interactions

Journal: International Journal of Biological Sciences

Article Title: Core transcriptional regulatory circuitry molecule ZNF217 promotes AML cell proliferation by up-regulating MYB

doi: 10.7150/ijbs.103211

Figure Lengend Snippet: MYB is identified as a key downstream target gene of ZNF217 in AML cells. A. WashU plot visual analysis highlights the binding sites (shaded) of ZNF217, ELF1, MEF2D, RUNX2, and FOXP1, while Hi-C analysis reveals their interaction relationships in MV4-11 cell line. B. In MV4-11 and U937 cells, interfering with ZNF217 resulted in decreased protein and mRNA expression of MYB. C. Knockout of the identified enhancer sequence led to decreased MYB protein expression. D. Knocking out the enhancer sequence in MV4-11 cas9 cells impaired cell proliferation. E. Results from the white slice experiment show that knocking out the enhancer sequence significantly decreased the MV4-11 cas9 cells proliferation in comparison to the control group. F. Knocking out the enhancer sequence in MV4-11 cells had no remarkable impact on cell proliferation.

Article Snippet: Next, we followed the flow cytometry intracellular staining protocol ( https://assets.thermofisher.com/TFS-Assets/BID/Technical-Notes/flow-intracellular-staining-en.pdf ) and used ZNF217 antibody along with FITC-conjugated secondary antibody (ThermoFisher, A-11034) to simultaneously detect ZNF217 protein expression levels in HSPCs.

Techniques: Binding Assay, Hi-C, Expressing, Knock-Out, Sequencing, Comparison, Control

Journal: International Journal of Biological Sciences

Article Title: Core transcriptional regulatory circuitry molecule ZNF217 promotes AML cell proliferation by up-regulating MYB

doi: 10.7150/ijbs.103211

Figure Lengend Snippet: Pattern diagram. A. ZNF217, ELF1, MEF2D, RUNX2 and FOXP1 collaboratively regulate ZNF217 expression by co-occupying the enhancer region of ZNF217. B. ZNF217, ELF1, MEF2D, RUNX2, FOPX1, and LDB1 engage in collaborative regulation of MYB transcription through the interaction between its promoter and the identified enhancer regions.

Article Snippet: Next, we followed the flow cytometry intracellular staining protocol ( https://assets.thermofisher.com/TFS-Assets/BID/Technical-Notes/flow-intracellular-staining-en.pdf ) and used ZNF217 antibody along with FITC-conjugated secondary antibody (ThermoFisher, A-11034) to simultaneously detect ZNF217 protein expression levels in HSPCs.

Techniques: Expressing

Journal: Journal of the Endocrine Society

Article Title: The PCOS GWAS Candidate Gene ZNF217 Influences Theca Cell Expression of DENND1A.V2 , CYP17A1 , and Androgen Production

doi: 10.1210/jendso/bvac078

Figure Lengend Snippet: A, ZNF217 immunohistochemical staining of normal ovarian tissue (right) and polycystic ovary syndrome (PCOS) ovarian tissue (left). Images were taken at 40× magnification. In comparison to the theca cells of the normal cycling ovary, ZNF217 protein expression was decreased in theca layer of follicles of PCOS ovarian specimens. Antibody neutralized with the ZNF217 immunogenic peptide was used in the bottom 2 images to confirm specificity of the immunoperoxidase signal. B, Comparison of ZNF217 staining intensity in the designated region of interest (ROI, theca interna) revealed a significant decrease in the theca cell layer of PCOS ovarian tissue (N = 4) compared to normal ovarian tissue (N = 4); * P < .001.

Article Snippet: Anti-ZNF217 antibody (Atlas Antibodies catalog No. HPA051857, RRID:AB_2681641, https://scicrunch.org/resources-legacy/Any/search?q=AB_2681641&l=AB_2681641 ), which binds to both ZNF217.V1 and ZNF217.V2), and the ZNF217 recombinant protein (APREST85710) were obtained from Sigma-Aldrich.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Journal of the Endocrine Society

Article Title: The PCOS GWAS Candidate Gene ZNF217 Influences Theca Cell Expression of DENND1A.V2 , CYP17A1 , and Androgen Production

doi: 10.1210/jendso/bvac078

Figure Lengend Snippet: ZNF217 expression in normal and polycystic ovary syndrome (PCOS) theca cells. A, ZNF217 (green signal) is localized in punctate nuclear structures in human theca cells by immunofluorescence. The ZNF217 signal is reduced in PCOS theca cells compared to normal theca cells cultured without forskolin, and in normal theca cells treated with forskolin (20 μM, 24 hours). B, Representative Western blots comparing ZNF217 and DENND1A.V2 protein expression in 2 normal and 2 PCOS theca cell preparations cultured in the C, absence, or F, presence of forskolin (20 μM) for 24 hours. C, Quantitative Western blot data from 5 normal and 5 PCOS preparations demonstrated that ZNF217 protein was decreased by forskolin treatment in normal theca cells ( α P < .001). ZNF217 protein in PCOS theca cells was decreased compared to normal theca cells (*** P < .001) under control culture conditions. D, Relative DENND1A.V2 protein normalized by mammalian target of rapamycin (mTOR) was elevated both in forskolin-treated (*** P < .005) and untreated (*** P < .005) PCOS cells.

Article Snippet: Anti-ZNF217 antibody (Atlas Antibodies catalog No. HPA051857, RRID:AB_2681641, https://scicrunch.org/resources-legacy/Any/search?q=AB_2681641&l=AB_2681641 ), which binds to both ZNF217.V1 and ZNF217.V2), and the ZNF217 recombinant protein (APREST85710) were obtained from Sigma-Aldrich.

Techniques: Expressing, Immunofluorescence, Cell Culture, Western Blot

Journal: Journal of the Endocrine Society

Article Title: The PCOS GWAS Candidate Gene ZNF217 Influences Theca Cell Expression of DENND1A.V2 , CYP17A1 , and Androgen Production

doi: 10.1210/jendso/bvac078

Figure Lengend Snippet: ZNF217 abundance in normal and polycystic ovary syndrome (PCOS) theca cells and the relationship between ZNF217 messenger RNA (mRNA) to dehydroepiandrosterone (DHEA) accumulation, DENND1A.V2, CYP17A1, and miR-130b-3p expression. Individual theca cell preparations from normal (n = 5) and PCOS (n = 5) women were cultured in the absence or presence of forskolin (20 μM). For quantitative reverse transcription polymerase chain reaction, mRNA was harvested from the cells 16 hours following treatment and ZNF217, CYP17A1, DENND1A.V2, miR-130b-3p expression was quantified. In parallel cultures of normal and PCOS theca cells, conditioned medium was collected at 48 hours and DHEA biosynthesis was assayed. A, PCOS theca cells showed decreased expression of ZNF217 mRNA compared to normal theca cells both in forskolin-treated (*** P < .001) and untreated cultures (** P < .001. In normal theca cells, the addition of forskolin significantly decreased ZNF217 mRNA (* P < .01). B, ZNF217 mRNA abundance compared to DHEA production. PCOS theca cells cultured in the absence or presence of forskolin showed increased DHEA production and decreased ZNF217 mRNA expression compared to normal theca cells. There was an inverse negative correlation between ZNF217 mRNA expression and DHEA production (Spearman ρ = –0.7353; P < .001). C, CYP17A1 gene expression in PCOS theca cells, required for androgen biosynthesis in PCOS, was negatively correlated with decreased ZNF217 mRNA abundance (Spearman ρ = –0.67890; P = .001). D, Comparison of ZNF217 mRNA expression to DENND1A.V2 mRNA abundance in PCOS and normal theca cells treated without or with forskolin revealed an inverse negative correlation (Spearman ρ = –0.6992; P < .001). E, miR-130b-3p, which targets DENND1A.V2 transcripts, was positively correlated with ZNF217 expression (Spearman ρ = 0.7815; P < .001).

Article Snippet: Anti-ZNF217 antibody (Atlas Antibodies catalog No. HPA051857, RRID:AB_2681641, https://scicrunch.org/resources-legacy/Any/search?q=AB_2681641&l=AB_2681641 ), which binds to both ZNF217.V1 and ZNF217.V2), and the ZNF217 recombinant protein (APREST85710) were obtained from Sigma-Aldrich.

Techniques: Expressing, Cell Culture, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of the Endocrine Society

Article Title: The PCOS GWAS Candidate Gene ZNF217 Influences Theca Cell Expression of DENND1A.V2 , CYP17A1 , and Androgen Production

doi: 10.1210/jendso/bvac078

Figure Lengend Snippet: Forced expression of ZNF217 in polycystic ovary syndrome (PCOS) theca cells converts PCOS theca cells to a normal phenotype of reduced DENND1A.V2, CYP17A1, and dehydroepiandrosterone (DHEA) biosynthesis using a ZNF217 adenovirus (Adv). PCOS theca cells (N=5) were infected with either 3 pfu/cell of Null (empty) vector adenovirus (Null Adv) or ZNF217 adenovirus (ZNF217 Adv) overnight, then treated in the absence (Control) and presence of forskolin (20 μM) for 36 hours for RNA studies or 72 hours for DHEA studies. A, ZNF217 Adv infection augmented ZNF217 messenger RNA (mRNA) expression under unstimulated (control, ***P < .0001) and forskolin-stimulated conditions ( **P < .001). ZNF217 Adv infection following forskolin stimulation resulted in decreased ZNF217 mRNA compared to control untreated cells ( α P < .01). B, Following ZNF217 Adv infection, PCOS theca cells were converted to a normal phenotype of diminished DENND1A.V2 mRNA accumulation, under both control (** P < .001) and forskolin-stimulated conditions (** P < .001), compared to Null Adv (control) infected cells. C, CYP17A1 mRNA accumulation was increased following forskolin-treatment ( α P < .01) in Null Adv–infected cells. In addition, ZNF217 Adv infection dramatically decreased CYP17A1 mRNA accumulation under both control (** P < .001) and forskolin-stimulated conditions (** P < .001). D, At 72 hours following infection with Null Adv, DHEA accumulation was observed to be increased by forskolin treatment ( α P < .01). ZNF217 Adv infection resulted in decreased DHEA biosynthesis under both control (** P < .001) and forskolin-stimulated cells (** P < .001). E, Forced ZNF217 overexpression with ZNF217 Adv, augmented miR-130b-3p expression under unstimulated (control, **P < .001) and forskolin-stimulated conditions ( *P < .1). ZNF217 Adv infection following forskolin stimulation resulted in decreased ZNF217 mRNA compared to control untreated cells ( α P < .01).

Article Snippet: Anti-ZNF217 antibody (Atlas Antibodies catalog No. HPA051857, RRID:AB_2681641, https://scicrunch.org/resources-legacy/Any/search?q=AB_2681641&l=AB_2681641 ), which binds to both ZNF217.V1 and ZNF217.V2), and the ZNF217 recombinant protein (APREST85710) were obtained from Sigma-Aldrich.

Techniques: Expressing, Infection, Plasmid Preparation, Over Expression

Journal: Journal of the Endocrine Society

Article Title: The PCOS GWAS Candidate Gene ZNF217 Influences Theca Cell Expression of DENND1A.V2 , CYP17A1 , and Androgen Production

doi: 10.1210/jendso/bvac078

Figure Lengend Snippet: ZNF217 knockdown in normal theca cells results in augmented DENND1A.V2 and CYP17A1 messenger RNA (mRNA) expression. Normal theca cells (N=5) were infected with equivalent amounts of control short hairpin RNA (shRNA) or ZNF217 shRNA lentiviral particles (3 pmol/10 6 cells) for 24 hours, then treated for 36 hours in the absence (control) or presence of 20 μM forskolin. A, In the absence of forskolin, ZNF217 expression was decreased following infection with ZNF217 lentiviral shRNA (* P < .05) as compared control shRNA. Forskolin treatment inhibited ZNF217 mRNA in control shRNA-infected cells ( α P < .05). B, DENND1A.V2 expression was increased following ZNF217 shRNA lentivirus infection under both control (*** P < .001) and forskolin-stimulated (*** P < .001) conditions. C, CYP17 A1 mRNA was increased by forskolin-stimulation in normal theca cells infected with control shRNA ( α P < .01). CYP17 A1 mRNA was increased following ZNF217 shRNA lentivirus infection under both control (* P < .01) and forskolin-stimulated (** P < .001) conditions, compared to control shRNA-infected cells.

Article Snippet: Anti-ZNF217 antibody (Atlas Antibodies catalog No. HPA051857, RRID:AB_2681641, https://scicrunch.org/resources-legacy/Any/search?q=AB_2681641&l=AB_2681641 ), which binds to both ZNF217.V1 and ZNF217.V2), and the ZNF217 recombinant protein (APREST85710) were obtained from Sigma-Aldrich.

Techniques: Expressing, Infection, shRNA

Journal: Journal of the Endocrine Society

Article Title: The PCOS GWAS Candidate Gene ZNF217 Influences Theca Cell Expression of DENND1A.V2 , CYP17A1 , and Androgen Production

doi: 10.1210/jendso/bvac078

Figure Lengend Snippet: A polycystic ovary syndrome (PCOS) genome-wide association study (GWAS) candidate gene network encompassing ZNF217 and DENND1A.V2. PCOS is a manifestation of altered expression of a network of PCOS GWAS candidate genes including LHCGR , ZNF217 , DENND1A , and RAB5B. Decreased ZNF217 expression in PCOS theca cells is associated with increased DENND1A.V2 and CYP17A1 expression, and downstream, androgen biosynthesis. Alternatively, ZNF217 could indirectly influence DENND1A.V2, RAB5B, and subsequently CYP17A1 expression, through the intermediacy of miR-130b-3p.

Article Snippet: Anti-ZNF217 antibody (Atlas Antibodies catalog No. HPA051857, RRID:AB_2681641, https://scicrunch.org/resources-legacy/Any/search?q=AB_2681641&l=AB_2681641 ), which binds to both ZNF217.V1 and ZNF217.V2), and the ZNF217 recombinant protein (APREST85710) were obtained from Sigma-Aldrich.

Techniques: GWAS, Expressing