Journal: Methods in Molecular Biology (Clifton, N.j.)

Article Title: Isolation of undifferentiated and early differentiating Type A spermatogonia from Pou5f1-GFP reporter mice

doi: 10.1007/978-1-61779-436-0_3

Figure Lengend Snippet: Undifferentiated and early differentiating spermatogonia isolated through Fluorescence Activated Cell Sorting (FACS). (A) Flow-cytometric analysis and sorting of wild-type and Pou5f1-GFPMann cells stained with APC-Kit antibody. (B) Real-time-PCR analysis of Kit and canonical markers of undifferentiated spermatogonia in Pou5f1+/Kit- and Pou5f1+/Kit+ cells sorted from A. Cells were processed for RT-PCR using the TaqMan Gene Expression Cells-to-CT Kit (Applied Biosystems, Carlsbad, CA) according to manufacturer’s instructions. The expression value of each gene was normalized to the amount of an internal control gene (Eif3land Rps3) (29) and a relative quantitative fold change was determined using the ΔΔ Ct method. Experiments were performed in triplicate and data are represented as mean ± standard error. Taqman Gene Expression Assays (Applied Biosystems, Carlsbad, CA) used for specific transcripts were: Mm00460859_m1 (Eif3l), Mm00833897_m1 (Gfra1), Mm00445212_m1 (Kit), Mm00437606_s1 (Neurog3), Mm00658129_gH (Pou5f1), Mm00656272_m1 (Rps3), Mm00493681_m1 (Thy1), and Mm01176868_m1 (Zbtb16). Normalization to Eif3l and Rps3 produced identical results.

Article Snippet: Taqman Gene Expression Assays (Applied Biosystems, Carlsbad, CA) used for specific transcripts were: Mm00460859_m1 ( Eif3l ), Mm00833897_m1 ( Gfra1 ), Mm00445212_m1 ( Kit ), Mm00437606_s1 ( Neurog3 ), Mm00658129_gH ( Pou5f1 ), Mm00656272_m1 ( Rps3 ), Mm00493681_m1 ( Thy1 ), and Mm01176868_m1 ( Zbtb16 ).

Techniques: Isolation, Fluorescence, FACS, Staining, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Expressing, Control, Produced

Journal: Communications biology

Article Title: PLZF and its fusion proteins are pomalidomide-dependent CRBN neosubstrates.

doi: 10.1038/s42003-021-02801-y

Figure Lengend Snippet: Fig. 3 G410 and G467 of PLZF are critical for its binding to CRBN in the presence of pomalidomide. a Schematic representation of the ZF motifs of PLZF. b Multiple sequence alignment showing PLZF ZF motifs along with validated ZF degrons from Ikaros, ZFP91, and SALL4. Cys and His residues (blue and green, respectively) comprising ZF motifs and critical Gly residues (red) are highlighted. c FLAG-HA (FH)-CRBN and Myc-FLAG (MF)-PLZF carrying point mutations were coexpressed in 293T cells and immunoprecipitated with anti-HA antibody in the presence or absence of pomalidomide. d Schematic representation of EGFP-fused PLZF ZF proteins. e FH-CRBN and EGFP-fused PLZF ZF proteins were coexpressed in 293T cells and immunoprecipitated with anti-Flag antibody in the presence or absence of pomalidomide. WCL whole-cell lysate. All experiments were conducted more than twice with similar results.

Article Snippet: Rabbit anti-human CRBN65 mAb (Celgene, San Diego, CA, 1:10000 dilution, Lot# CGN-6-4-5), mouse anti-PLZF mAb (39987, Active Motif, 1:1000 dilution, Lot#05313002), mouse anti-PLZF mAb (sc-28319, Santa Cruz, 1:1000 dilution, Lot#C0618), mouse anti-SALL4 mAb (ab57577, Abcam, 1:1000 dilution, Lot#GR3198290-2), mouse anti-Vinculin mAb (ab18058, Abcam, 1:1000 dilution, Lot#GR207163-1), mouse anti-HA11 mAb (901503, BioLegend, 1:1000 dilution, Lot#b207273), mouse anti-Flag M2 mAb (F1804, SIGMA, 1:1000 dilution, Lot#SLBS3530V), rabbit anti-Myc tag (ab9106, Abcam, 1:1000 dilution), rabbit anti-GSPT1 (ab49878, Abcam, 1:1000 dilution, Lot#GR274469-4), rabbit antiCK1α mAb (ab108296, Abcam, 1:1000 dilution, Lot#GR53415-9), and rabbit antiIkaros mAb (ab26083, Abcam, 1:1000 dilution, Lot#GR250435-1), rabbit anti-GFP (ab290, Abcam, 1:1000 dilution, Lot#GR3184825-1) were used as primary antibodies.

Techniques: Binding Assay, Sequencing, Immunoprecipitation

Journal: Molecular & Cellular Proteomics : MCP

Article Title: The Glial Cell-Derived Neurotrophic Factor (GDNF)-responsive Phosphoprotein Landscape Identifies Raptor Phosphorylation Required for Spermatogonial Progenitor Cell Proliferation *

doi: 10.1074/mcp.M116.065797

Figure Lengend Snippet: GDNF overexpression results in mTORC1 activation in SPCs in vivo. A, Tubules from testes overproducing GDNF after transduction with a GDNF-encoding lentivirus (GDNF OE) contained clumps of spermatogonia-like cells. Positivity for the marker ZBTB16 confirmed that these cells were spermatogonia. Accumulating spermatogonia in GDNF-overexpressing testis were positive for phosphorylated ERK1/2, which is normally restricted to Sertoli cells. Accumulated spermatogonia also exhibited high mTORC1 activity reflected by phosphorylated RPS6, particularly in cells at the basement membrane of seminiferous tubules. Scale bar, 20 μm. B, Western blotting analyses of two independent samples and control sets demonstrated elevated levels of phosphorylated ERK1/2, RPS6, and raptor Ser863 in GDNF overexpressing testes compared with normal testes, which were likely contributed by accumulated SPCs.

Article Snippet: Immunostaining of sections was performed for ZBTB16 (Cat#AF2944, R&D Systems) ( 42 , 43 ), LIN28A (Cat#ab4602, Abcam, Shanghai, China) ( 44 ) phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cat#9106, Cell Signaling Technology), and phospho-S6 Ribosomal Protein (Ser240/244) (Cat#5364, Cell Signaling Technology).

Techniques: Over Expression, Activation Assay, In Vivo, Transduction, Marker, Activity Assay, Western Blot

Journal: Journal of Cellular Biochemistry

Article Title: ZBTB16 as a Downstream Target Gene of Osterix Regulates Osteoblastogenesis of Human Multipotent Mesenchymal Stromal Cells

doi: 10.1002/jcb.25634

Figure Lengend Snippet: ZBTB16 expression depends on OIM during osteoblastic differentiation. (A) Three hMSCs (hMSCs #1, #2, and #3) were cultured with or without OIM (containing AA, β‐GP, and Dex) for 4, 7, and 14 d. The mRNA levels of ZBTB16 were evaluated by TaqMan assays. N.D.: not detectable. The bars represent the mean ± S.D. of triplicate wells. The results are representative of three independent experiments. β‐Actin was used as an internal control. (B) hBMMSCs were cultured with or without OIM for 4, 7, and 14 d. (C) Western blot analysis of ZBTB16 using cell lysates from hMSCs cultured with or without OIM for 4, 7, and 14 d. β‐Actin was used as an internal control. Representative data from three different samples are shown in (B) and (C); a similar tendency was observed in other samples. The experiments shown in Figure A and B were performed in triplicate.

Article Snippet: The primers used were as follows: ZBTB16 (Hs00957433_m1), Osx (Hs00541729_m1), RUNX2 (Hs00231692_m1), OCN (Hs01587814_g1), BSP (Hs00173720_m1), and β‐actin (43263251E; all of the probes were obtained from Applied Biosystems).

Techniques: Expressing, Cell Culture, Control, Western Blot

Journal: Journal of Cellular Biochemistry

Article Title: ZBTB16 as a Downstream Target Gene of Osterix Regulates Osteoblastogenesis of Human Multipotent Mesenchymal Stromal Cells

doi: 10.1002/jcb.25634

Figure Lengend Snippet: The inhibitory effect of siZBTB16 on ALP activity, mineralized nodule formation, and osteogenic gene expression in hMSCs. (A) The efficacy of the siRNA inhibiting ZBTB16 mRNA expression. (B) The effect of siRNA targeting ZBTB16 in hMSCs on the osteoblastic differentiation of hMSCs. The ALP activity of hMSCs after transfection and culture with or without OIM for 5 d. (C) The effect of siRNA targeting ZBTB16 on the osteoblastic differentiation of hBMMSCs. The ALP activity of hBMMSCs after they were transfected and cultured with or without OIM for 5 d. (D) The cell viability of hMSCs was measured by MTS assay after they were transfected and cultured with or without OIM for 5 d. * P < 0.05, compared with non‐siRNA and siControl. (E) Alizarin red S staining of hMSCs after the cells were transfected with siZBTB16 and cultured with or without OIM for 21 d. (F) Bound stain was eluted with a solution of 10% cetylpyridinium chloride and quantified by measuring the absorbance at a wavelength of 570 nm using a plate reader. (G) Effect of a siRNA targeting ZBTB16 in hMSCs on OCN and BSP expression after hMSCs were transfected and cultured with OIM for 5 d. * P < 0.05, compared with siControl. (−), non‐siRNA; siCont., siControl. These figures show representative data from three different samples; a similar tendency was observed in other hMSCs. The experiments shown in Figure A, B–D, F, and G were performed in triplicate.

Article Snippet: The primers used were as follows: ZBTB16 (Hs00957433_m1), Osx (Hs00541729_m1), RUNX2 (Hs00231692_m1), OCN (Hs01587814_g1), BSP (Hs00173720_m1), and β‐actin (43263251E; all of the probes were obtained from Applied Biosystems).

Techniques: Activity Assay, Gene Expression, Expressing, Transfection, Cell Culture, MTS Assay, Staining

Journal: Journal of Cellular Biochemistry

Article Title: ZBTB16 as a Downstream Target Gene of Osterix Regulates Osteoblastogenesis of Human Multipotent Mesenchymal Stromal Cells

doi: 10.1002/jcb.25634

Figure Lengend Snippet: The mRNA and protein levels of ZBTB16 were decreased by siOsx. (A) hMSCs were transfected with non (siCont.)‐ or Osx (siOsx1 and siOsx2)‐specific siRNA and cultured with OIM. RNA was extracted, and real‐time RT‐PCR was performed for Osx and ZBTB16. (B) hMSCs were transfected with non (siCont.)‐ or RUNX2 (siRUNX2)‐specific siRNA and cultured with OIM. RNA was extracted, and real‐time RT‐PCR was performed for RUNX2 and ZBTB16. (C) Western blot analysis of ZBTB16 using cell lysates from hMSCs after they were transfected and cultured with OIM. * P < 0.05, compared with siControl. These figures show representative data from three different samples; a similar tendency was observed in other hMSCs. The experiments shown in Figure A and B were performed in triplicate.

Article Snippet: The primers used were as follows: ZBTB16 (Hs00957433_m1), Osx (Hs00541729_m1), RUNX2 (Hs00231692_m1), OCN (Hs01587814_g1), BSP (Hs00173720_m1), and β‐actin (43263251E; all of the probes were obtained from Applied Biosystems).

Techniques: Transfection, Cell Culture, Quantitative RT-PCR, Western Blot

Journal: Journal of Cellular Biochemistry

Article Title: ZBTB16 as a Downstream Target Gene of Osterix Regulates Osteoblastogenesis of Human Multipotent Mesenchymal Stromal Cells

doi: 10.1002/jcb.25634

Figure Lengend Snippet: Osx binds to the Sp1 sequence of the ZBTB16 promoter region. (A) The diagram shows the location of the primers used to amplify the Sp1 or the non‐Sp1 site of the ZBTB16 promoter region in the ChIP assay. TSS, transcription start site of ZBTB16; P1, primer 1 set; P2, primer 2 set; P3 (negative control), primer 3 set. (B) hMSCs were cultured with OIM and rhBMP‐6 (200 ng/ml) for 7 d. ChIP assay was performed with antibodies against Osx and IgG (negative control). Ten percent of the input was loaded as a control. This figure shows representative data from three different samples; a similar tendency was observed in other hMSCs.

Article Snippet: The primers used were as follows: ZBTB16 (Hs00957433_m1), Osx (Hs00541729_m1), RUNX2 (Hs00231692_m1), OCN (Hs01587814_g1), BSP (Hs00173720_m1), and β‐actin (43263251E; all of the probes were obtained from Applied Biosystems).

Techniques: Sequencing, Negative Control, Cell Culture, Control

Journal: Cell

Article Title: RAB11FIP5 Expression and Altered Natural Killer Cell Function Are Associated with Induction of HIV Broadly Neutralizing Antibody Responses

doi: 10.1016/j.cell.2018.08.064

Figure Lengend Snippet:

Article Snippet: Human PLZF Allophycocyanin mAb (Clone 6318100) antibody , R and D Systems , Cat# IC2944A; RRID: AB_10730709.

Techniques: Recombinant, Staining, Cell Isolation, Enzyme-linked Immunosorbent Assay, Marker, Library Quantification, Gene Expression, Sequencing, Software

Journal: bioRxiv

Article Title: Differential retinoic acid responses across testicular development in vitro

doi: 10.64898/2025.12.30.696417

Figure Lengend Snippet: CIVMs derived from neonatal testis from PND 5 and PND 10 testis were exposure to isotretinoin for 24 hours. (A) Expression of Stra8 in PND 5 derived CIVMs determined by RT-qPCR. In the control, 0.3, 3, and 30 nM isotretinoin dose groups the expression of Stra8 contained samples below the threshold of quantification. The expression of Stra8 was significantly increased in the 300 nM isotretinoin dose group (p-value <0.0001). (B) Expression of Plzf in PND 5 derived CIVMs determined by RT-qPCR. All samples were above the threshold of quantification. The expression of Plzf in the 300 nM isotretinoin dose group was significantly lower than control (p-value of 0.026). (C) Expression of Stra8 in PND 10 derived CIVMs determined by RT-qPCR. Aside from one control sample, all samples were above the threshold of quantification. The expression of Stra8 was significantly increased in the 3,30, and 300 nM isotretinoin dose groups (p-values all <0.0001). (D) Expression of Plzf in PND 10 derived CIVMs determined by RT-qPCR. All samples were above the threshold of quantification. The expression of Plzf in the 30 and 300nM isotretinoin dose groups were significantly lower than control (p-values <0.0001).

Article Snippet: RT-qPCR was used to target the rat genes Stra8 (Rn01747849_m1) and Plzf (Rn01418644_m1).

Techniques: Derivative Assay, Expressing, Quantitative RT-PCR, Control

Journal: bioRxiv

Article Title: Differential retinoic acid responses across testicular development in vitro

doi: 10.64898/2025.12.30.696417

Figure Lengend Snippet: The expression of Plzf and Stra8 quantified by RT-qPCR at day in vitro 4 and day in vitro 15 in PND 5 tissue CIVMs and PND 10 tissue CIVMs in control and media supplementation conditions. Color indicates day in vitro , dashed line divides plot by PND 5 CIVMs and PND 10 CIVMs. Points that are “X” indicate samples that were below the limit of quantification (cycle threshold of 37). Data displayed as expression relative to beta-actin (housekeeping gene (HK), (2^ [gene – HK])).

Article Snippet: RT-qPCR was used to target the rat genes Stra8 (Rn01747849_m1) and Plzf (Rn01418644_m1).

Techniques: Expressing, Quantitative RT-PCR, In Vitro, Control

Journal: PLoS ONE

Article Title: Tzfp Represses the Androgen Receptor in Mouse Testis

doi: 10.1371/journal.pone.0062314

Figure Lengend Snippet: (A) Overview of the Tzfp protein with the BTB/POZ and zink finger domains (top), and of the wild-type and mutated Tzfp allele upon insertional mutagenesis (middle). The wild-type and mutated Tzfp transcript is also included (bottom). (B) Genotyping gel showing fragments indicating samples from wild-type, heterozygous and Tzfp GTi/GTi mice. (C) Gene expression analysis of Mll4 and Plzf in wild-type and Tzfp GTi/GTi testis reveals no significant difference between wild-type and Tzfp GTi/GTi testis. (D) Crosses between heterozygous animals reveal a sex-ratio distortion and fewer than expected homozygous mutant pups. The χ2 -test was used to determine significance. (E) Weight distribution of testis and epididymus in wild-type and Tzfp GTi/GTi mouse reveal a small, but not significant reduction in testis weight in the Tzfp GTi/GTi mice.

Article Snippet: Plzf: Mm01176865_m1.

Techniques: Mutagenesis, Gene Expression

Journal: iScience

Article Title: Glucocorticoid-mediated induction of ZBTB16 affects insulin secretion in human islets and EndoC-βH1 β-cells

doi: 10.1016/j.isci.2023.106555

Figure Lengend Snippet: ZBTB16 is the most strongly predicted direct glucocorticoid target in human islets and β-cells (A) Schematic summary of the bioinformatics workflow to rank differentially expressed genes according to their potential of being direct Glucocorticoid Receptor targets. (B) Top 10 differentially expressed genes upon 2 μM dexamethasone treatment with the lowest rank product. The rank product can be interpreted as a p-value, which represents the probability of the gene to be a direct target of GR. (C) Volcano plot representing differentially expressed genes upon 2 μΜ dexamethasone treatment and ZBTB16 ’s relative position compared to all genes. Significantly downregulated genes are indicated in red on the left side of the vertical dashed line and significantly upregulated genes are indicated in red on the right of the vertical dashed line. Vertical dashed lines correspond to the log2 fold change threshold of 0 separating down- and upregulation and horizontal dashed lines correspond to the adjusted p-value threshold of 0.05 represented as log 10 adjusted p-value. (D) ZBTB16 RNA expression (RNA-seq, left, n = 4 biological replicates) and protein expression (Western blot, right, n = 6 biological replicates) in EndoC-βΗ1 after 2 μM dexamethasone treatment. Ctrl; Control (DMSO), Dexa; Dexamethasone. Data are presented as mean ± SEM; ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: ZBTB16 (PLZF) (D8G3G) Rabbit mAb , Cell Signaling Technology , #39784.

Techniques: RNA Expression, RNA Sequencing, Expressing, Western Blot, Control

Journal: iScience

Article Title: Glucocorticoid-mediated induction of ZBTB16 affects insulin secretion in human islets and EndoC-βH1 β-cells

doi: 10.1016/j.isci.2023.106555

Figure Lengend Snippet: Glucocorticoid responsive elements (GREs) are located in intronic and conserved regions of ZBTB16 in human β-cells (A) Distribution of predicted GREs across ZBTB16 gene on the human islets. (B) Predicted GRE overlaps (and their genomic coordinates) visualized with regulatory/conservation tracks from UCSC Genome Browser; tracks represent from top to bottom: ZBTB16 transcript localization, ENCODE regulatory elements, GeneHancer regulatory elements, Open chromatin positions (DNase I peak clusters), Phylop basewise evolutionary/conservation scores. Red lines on the peak boxes (positions 5, 6, 8, and 10) correspond to GR binding sequence (GBS) occurrences. (C) Glucocorticoid Receptor ChIP followed by PCR for the predicted GRE coordinates. Position numbers correspond to the positions of (A) from left to right (see also Figure S1 ). Ctrl; Control (DMSO), Dexa; Dexamethasone (100 nM).

Article Snippet: ZBTB16 (PLZF) (D8G3G) Rabbit mAb , Cell Signaling Technology , #39784.

Techniques: Binding Assay, Sequencing, Control

Journal: iScience

Article Title: Glucocorticoid-mediated induction of ZBTB16 affects insulin secretion in human islets and EndoC-βH1 β-cells

doi: 10.1016/j.isci.2023.106555

Figure Lengend Snippet: Dexamethasone induces ZBTB16 expression in a dose- and time-dependent manner and impairs insulin secretion in EndoC-βH1 cells (A) Dose-response curve of increasing concentration of dexamethasone treatment (0.1, 0.5, 1, 10, 100, 1000, and 2000 nM) for 24h in the expression levels of ZBTB16 , SGK1, and GR (n = 4 biological replicates). Black stars indicate gene-specific expression differences between successive doses (e.g., 1 vs. 10, 10 vs. 100, 100 vs. 1000, and 1000 vs. 2000 nM). Red stars illustrate expression differences between ZBTB16 and SGK1 . ZBTB16 and SGK1 expression levels are significantly different from that of GR in all treatment doses (significance symbols omitted for clarity). (B) Cell viability (MTS) assay relative to the control after treatment with increasing dexamethasone concentrations (n = 4 biological replicates for 100 nM, 3 biological replicates for 1 and 2 μΜ) for 48h. (C) Gene expression levels of ZBTB16 , SGK1, and GR upon treatment with 100 nM dexamethasone at different time points (2, 8, 24, and 48h) (n = 4 biological replicates). The black star indicates gene-specific expression difference at each time point compared to 2h post-treatment. Red stars illustrate expression differences between ZBTB16 and SGK1 . ZBTB16 and SGK1 expression levels are significantly different from that of GR in all time points (significance symbols omitted for clarity). (D) Insulin secretion of EndoC-βH1 cells upon treatment with 100 nM dexamethasone at different time points (n = 6 biological replicates). All comparisons between low and high glucose treatments were significantly different regardless of treatment but the symbols for significance were omitted for clarity. (E) Insulin content of EndoC-βH1 cells upon treatment with 100 nM dexamethasone at different time points (n = 6 biological replicates). Forevery grouped comparison repeated measures ANOVA (1-, 2-, or 3-way) was performed. Post-hoc pairwise comparisons included Tukey multiple comparisons for cell viability, dose- and time-dependent dexamethasone treatment experiments and paired t-tests for insulin secretion and insulin content experiments. Low glucose; 1 mM, high glucose; 20 mM, Ctrl; Control (DMSO), Dexa; Dexamethasone. Data are presented as mean ± SEM; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: ZBTB16 (PLZF) (D8G3G) Rabbit mAb , Cell Signaling Technology , #39784.

Techniques: Expressing, Concentration Assay, MTS Assay, Control, Gene Expression, Comparison

Journal: iScience

Article Title: Glucocorticoid-mediated induction of ZBTB16 affects insulin secretion in human islets and EndoC-βH1 β-cells

doi: 10.1016/j.isci.2023.106555

Figure Lengend Snippet: ZBTB16 overexpression and induction suppression affect gene expression and insulin secretion in EndoC-βΗ1 cells (A) Relative expression of ZBTB16 (compared to Ctrl) after ZBTB16 overexpression as measured by qPCR assay. (B) Expression induction of ZBTB16 (fold-change relative expression; Dexa/Ctrl expression) in control and dexamethasone-treated samples after ZBTB16 overexpression. (C) Insulin secretion (left) and insulin content (right) after ZBTB16 overexpression. (D) Relative expression of SGK1 , (E) GR , (F) PDX-1 , (G) INS (compared to Ctrl) after ZBTB16 overexpression as measured by qPCR assay. (H) Relative expression of ZBTB16 (compared to NC) after ZBTB16 induction suppression as measured by qPCR assay. (I) Expression induction of ZBTB16 (fold-change relative expression; Dexa/Ctrl expression) in NC and dexamethasone-treated samples after ZBTB16 induction suppression. (J) Insulin secretion (left) and insulin content (right) after ZBTB16 induction suppression. (K) Relative expression of SGK1 , (L) GR , (M) PDX-1 , (N) INS (compared to NC) after ZBTB16 induction suppression as measured by qPCR assay. All experiments were conducted on EndoC-βH1 cells (n = 5–6 biological replicates). Comparisons between qPCR results were performed using the Wilcoxon matched pairs signed rank test. Insulin secretion and content grouped comparisons were performed with repeated measures two-way ANOVA followed by pairwise comparisons with paired Student’s t test. Low glucose; 1 mM, high glucose; 20 mM, Ctrl; Control (DMSO), Dexa; Dexamethasone (100 nM), pcControl; Control plasmid pcDNA3.1, ZBTB16_OE; ZBTB16 overexpression, NC; Negative Control, ZBTB16_KD; ZBTB16 knockdown. Data are presented as mean ± SEM; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: ZBTB16 (PLZF) (D8G3G) Rabbit mAb , Cell Signaling Technology , #39784.

Techniques: Over Expression, Gene Expression, Expressing, Control, Plasmid Preparation, Negative Control, Knockdown

Journal: iScience

Article Title: Glucocorticoid-mediated induction of ZBTB16 affects insulin secretion in human islets and EndoC-βH1 β-cells

doi: 10.1016/j.isci.2023.106555

Figure Lengend Snippet: ZBTB16 predicted direct gene targets are involved in several regulatory pathways, including mitochondrial function (A) Schematic summary of the bioinformatics workflow to identify ZBTB16 predicted direct gene targets in human pancreatic islets. (B) Subset of potential islet ZBTB16 targets expressed in human pancreatic β-cells. (C) Bar chart showing selected enriched terms/pathways derived from the functional annotation of the ZBTB16 predicted direct gene targets in human islets ( Up ) and human β-cells ( Down ). (D) Mitochondrial oxygen consumption measurements in EndoC-βH1 cells (n = 6 biological replicates) corresponding to cellular respiration levels comparing control cells (pcControl + Ctrl), control cells under dexamethasone treatment (pcControl + Dexa) and cells with overexpression of ZBTB16 under dexamethasone treatment (ZBTB16_OE + Dexa), (E) Quantification of baseline OCR levels, (F) Acute response measured as increase in OCR upon pyruvate injection, (G) Maximal respiration measured after addition of an uncoupler of the inner mitochondrial membrane (FCCP), (H) ATP production assessed as decrease in OCR after oligomycin injection, which inhibits ATP-synthase and (I) Coupling efficiency calculated as ATP production in relation to combined baseline and pyruvate response. Comparisons between conditions in each measurement were performed with one-way ANOVA followed by pairwise comparisons with Tukey’s multiple comparison test. pcControl; Control plasmid pcDNA3.1, ZBTB16_OE; ZBTB16 overexpression, Ctrl; Control (DMSO), Dexa; Dexamethasone (100 nM), FCCP; carbonyl cyanide p-trifluoro-methoxyphenyl hydrazone. Data are presented as mean ± SEM; ∗p < 0.05.

Article Snippet: ZBTB16 (PLZF) (D8G3G) Rabbit mAb , Cell Signaling Technology , #39784.

Techniques: Derivative Assay, Functional Assay, Control, Over Expression, Injection, Membrane, Comparison, Plasmid Preparation

Journal: iScience

Article Title: Glucocorticoid-mediated induction of ZBTB16 affects insulin secretion in human islets and EndoC-βH1 β-cells

doi: 10.1016/j.isci.2023.106555

Figure Lengend Snippet:

Article Snippet: ZBTB16 (PLZF) (D8G3G) Rabbit mAb , Cell Signaling Technology , #39784.

Techniques: Transplantation Assay, Recombinant, Gene Expression, Staining, Western Blot, Transfection, Isolation, Reverse Transcription, Bicinchoninic Acid Protein Assay, Enzyme-linked Immunosorbent Assay, Proliferation Assay, RNA Sequencing, Binding Assay, Negative Control, Software