Journal: Human Genome Variation

Article Title: Germline and somatic genetic changes in multicentric tumors obtained from a patient with multiple endocrine neoplasia type 1

doi: 10.1038/hgv.2017.13

Figure Lengend Snippet: Clinical and pathological manifestations in this MEN1 patient. CT image and pathology specimens. The patient had more than 10 tumors in the pancreas and the duodenum. The pancreas head had one tumor (7 mm in diameter), the body had two tumors (5, 3 mm) and the tail had two tumors (19, 5 mm). In the duodenum, the bulb had one tumor (5 mm), and the descending limb had one tumor (10 mm) and smaller ones (about 1 mm). ( a ) Duodenum. ( b ) Pancreatic head. ( c ) Pancreatic body. ( d ) Pancreatic tail. Yellow arrows indicate tumor. ( e ) Pedigree of the patient. P indicates the proband and E+ indicates germline mutation of the MEN1 gene (c.1505dupA). CT, computed tomography; MEN1, multiple endocrine neoplasia type 1.

Article Snippet: TaqMan primers/probes Hs00270415_cn (Chr11:64577357 on build GRCh37) (Probe1) and Hs01778293_cn (Chr11:64571825 on build GRCh37) (Probe2) (Thermo Fisher Scientific) were used for the analyses.

Techniques: Mutagenesis, Computed Tomography

Journal: Human Genome Variation

Article Title: Germline and somatic genetic changes in multicentric tumors obtained from a patient with multiple endocrine neoplasia type 1

doi: 10.1038/hgv.2017.13

Figure Lengend Snippet: Whole-exome sequencing of the MEN1 gene of a patient with MEN1. ( a ) IGV analysis, ( b ) sanger sequencing results and ( c ) pyrosequencing results. IGV, Integrative Genomics Viewer; MEN1, multiple endocrine neoplasia type 1.

Article Snippet: TaqMan primers/probes Hs00270415_cn (Chr11:64577357 on build GRCh37) (Probe1) and Hs01778293_cn (Chr11:64571825 on build GRCh37) (Probe2) (Thermo Fisher Scientific) were used for the analyses.

Techniques: Sequencing

Journal: Human Genome Variation

Article Title: Germline and somatic genetic changes in multicentric tumors obtained from a patient with multiple endocrine neoplasia type 1

doi: 10.1038/hgv.2017.13

Figure Lengend Snippet: Genotype of the MEN1 gene and somatic mutations

Article Snippet: TaqMan primers/probes Hs00270415_cn (Chr11:64577357 on build GRCh37) (Probe1) and Hs01778293_cn (Chr11:64571825 on build GRCh37) (Probe2) (Thermo Fisher Scientific) were used for the analyses.

Techniques: Mutagenesis, Sequencing

Journal: Human Genome Variation

Article Title: Germline and somatic genetic changes in multicentric tumors obtained from a patient with multiple endocrine neoplasia type 1

doi: 10.1038/hgv.2017.13

Figure Lengend Snippet: Copy number variation of the MEN1 gene of a MEN1 Patient. ( a ) Position of probes and mutation on the MEN1 gene. ( b ) Copy number analysis of the MEN1 gene by digital or real-time PCR analysis.

Article Snippet: TaqMan primers/probes Hs00270415_cn (Chr11:64577357 on build GRCh37) (Probe1) and Hs01778293_cn (Chr11:64571825 on build GRCh37) (Probe2) (Thermo Fisher Scientific) were used for the analyses.

Techniques: Mutagenesis, Real-time Polymerase Chain Reaction

Journal: Human Genome Variation

Article Title: Germline and somatic genetic changes in multicentric tumors obtained from a patient with multiple endocrine neoplasia type 1

doi: 10.1038/hgv.2017.13

Figure Lengend Snippet: Immunohistochemical staining of menin for normal and tumor tissues of a patient with MEN1. ( a ) Menin staining in the pancreas using an N-terminal menin antibody. ( b ) Menin staining in the pancreas using a C-terminal menin antibody. Tumor can be seen on the right side of the micrograph, while normal tissue can be seen on the left side. ( c ) Menin staining in the duodenum using an N-terminal menin antibody. ( d ) Menin staining in the duodenum using a C-terminal antibody (both micrographs show tumor tissues). MEN1, multiple endocrine neoplasia type 1.

Article Snippet: TaqMan primers/probes Hs00270415_cn (Chr11:64577357 on build GRCh37) (Probe1) and Hs01778293_cn (Chr11:64571825 on build GRCh37) (Probe2) (Thermo Fisher Scientific) were used for the analyses.

Techniques: Immunohistochemical staining, Staining

Journal: Oncotarget

Article Title: Menin mediates Tat-induced neuronal apoptosis in brain frontal cortex of SIV-infected macaques and in Tat-treated cells

doi: 10.18632/oncotarget.14993

Figure Lengend Snippet: Menin expression is visible in nuclei, with some cytoplasmic expression, in the frontal cortex of SIV-infected A . and control B . macaques. There are more menin-positive cells in SIV-infected macaques A, C . than in the controls B, D . IOD analysis shows more menin immunostaining in the frontal cortex of SIV-infected macaques ( #1-#9 ) compared with control macaques ( #10–13 ), integrated optical density = optical density - background E . data are expressed as mean ± SD, * P < 0.05. Western blotting also shows increased menin expression in SHIV -SF162.P4 -infected macaques ( #1 ) compared with control macaques ( #12 ). A specific β-actin band (about 43 kDa) is shown under the menin band (about 68 kDa) F . Original magnification: (A-B) 200×; (C, D) 400×. (A, C) from macaque #9 , (B, D) from #10 . Arrows showing positive cells of IHC staining.

Article Snippet: We also performed double-labeled IHC against menin (1:200, rabbit, Cell Signaling Technology, MA, USA, #6891) using DAB/peroxidase and GFAP (1:200, Epitomics, Burlingame, CA, USA, 2301-1) with Vector blue/alkaline phosphatase using the same method.

Techniques: Expressing, Infection, Immunostaining, Western Blot, Immunohistochemistry

Journal: Oncotarget

Article Title: Menin mediates Tat-induced neuronal apoptosis in brain frontal cortex of SIV-infected macaques and in Tat-treated cells

doi: 10.18632/oncotarget.14993

Figure Lengend Snippet: Menin expression (dark blue) is mainly observed in the nuclei of neurons (NeuN brown) in the frontal cortex of SIV-infected macaques A . Menin (dark blue) is also positive in activated microglial cytoplasm and processes, but not in microglial nuclei (Iba1, red) B . Menin (brown) is rarely stained in astrocytes of the cerebral cortex (GFAP, dark blue) C . but is positive in membranes and processes of white matter astrocytes (GFAP, dark blue) D . Representative double-labeled immunofluorescence images show NeuN (green) and menin (red) expression in the frontal cortex E–F . Menin expression is increased in SIV-infected macaques (E) compared with control macaques (F). Analysis of IOD of the double-positive area (yellow) shows significantly increased menin expression in neuronal nuclei of the SIV-infected macaques ( #2, 4, 7, 8 ) compared with control macaques ( #10–13 ) G . Data are expressed as mean ± SD, * P < 0.05. Original magnification: (A–D) 800×, (E–F) 400×. (A) From macaque #7 ; (B) from macaque #1; (C, D) from macaque #3 . (E) from macaque #7 ; (F) from macaque #13 . Arrows showing positive cells of double IHC staining.

Article Snippet: We also performed double-labeled IHC against menin (1:200, rabbit, Cell Signaling Technology, MA, USA, #6891) using DAB/peroxidase and GFAP (1:200, Epitomics, Burlingame, CA, USA, 2301-1) with Vector blue/alkaline phosphatase using the same method.

Techniques: Expressing, Infection, Staining, Labeling, Immunofluorescence, Immunohistochemistry

Journal: Oncotarget

Article Title: Menin mediates Tat-induced neuronal apoptosis in brain frontal cortex of SIV-infected macaques and in Tat-treated cells

doi: 10.18632/oncotarget.14993

Figure Lengend Snippet: Astrocytic gliosis is shown in the frontal cortex with GFAP IHC A . TUNEL-positive cells B . and ssDNA-positive cells C . are mainly small neuronal and glial cells. Arrows showing positive cells of IHC staining (A, B, C). Apoptosis of pyramidal neurons and small neurons of the cortex is seen by double-labeled IHC for ssDNA (blue) and NeuN (brown, D ). Menin- (brown) and cleaved-caspase 3- (dark blue) double-labeled cells E . Arrows showing positive cells of double IHC staining D, E . Significantly positive correlation ( P = 0.0118, R = 0.6722) is demonstrated between the number of cleaved-caspase 3-positive cells and menin-positive cells in 13 macaques F . There is a significant negative correlation between the number of NeuN-positive cells and menin-positive cells in 13 macaques ( P = 0.0069, R = −0.707) G . Original magnification: (A–E) 400×. (A) From macaque #9 ; (B-C) from macaque #6 ; (D) from #5 ; (E) from #4 .

Article Snippet: We also performed double-labeled IHC against menin (1:200, rabbit, Cell Signaling Technology, MA, USA, #6891) using DAB/peroxidase and GFAP (1:200, Epitomics, Burlingame, CA, USA, 2301-1) with Vector blue/alkaline phosphatase using the same method.

Techniques: TUNEL Assay, Immunohistochemistry, Labeling

Journal: Oncotarget

Article Title: Menin mediates Tat-induced neuronal apoptosis in brain frontal cortex of SIV-infected macaques and in Tat-treated cells

doi: 10.18632/oncotarget.14993

Figure Lengend Snippet: SH-SY5Y cells were transfected with pRK5M-Tat-flag or pRK5M-flag for 24 hours and subjected to IHC staining. Menin (green) and Tat (red) co-localized in the nuclei of SH-SY5Y cells A . Optical density assay showed that menin expression is significantly increased in SH-SY5Y cells transfected with pRK5M-Tat-flag compared with the group transfected with pRK5M-flag B . Representative western blot were shown C . Optical density analysis of menin expression D . Optical density analysis of cleaved caspase3 expression E . Primary neurons were treated with or without Tat (100 ng/mL) for 48 h. Menin and cleaved-caspase 3 is significantly increased in Tat-treated neurons compared with controls F-H . TUNEL staining shows significantly increased apoptosis (green) in pRK5M-Tat-flag-transfected SH-SY5Y cells I-J . Data are plotted as mean ± SD (n = 3). * P < 0.05, ** P < 0.01.

Article Snippet: We also performed double-labeled IHC against menin (1:200, rabbit, Cell Signaling Technology, MA, USA, #6891) using DAB/peroxidase and GFAP (1:200, Epitomics, Burlingame, CA, USA, 2301-1) with Vector blue/alkaline phosphatase using the same method.

Techniques: Transfection, Immunohistochemistry, Expressing, Western Blot, TUNEL Assay, Staining

Journal: The American Journal of Pathology

Article Title: miR-24 Inhibition Increases Menin Expression and Decreases Cholangiocarcinoma Proliferation

doi: 10.1016/j.ajpath.2016.10.021

Figure Lengend Snippet: Menin is down-regulated in CCA. A and B: By real-time PCR and immunoblots, menin expression is decreased in CCA cell lines compared to H69. Significance is shown versus H69 cells. C: Flow cytometry analysis demonstrated a decrease in menin protein expression in Mz-ChA-1 cells compared to H69 cells. D: By real-time PCR, menin expression decreased in advanced-stage human CCA tissue biopsy specimens compared with normal control. Data are expressed as means ± SEM performed in triplicate (A–D). n = 3 independent samples (A and B). ∗P < 0.05 versus H69 or human control. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Modulation of Menin Expression, Proliferation, and Angiogenesis in Vitro Mz-ChA-1 cells were targeted for the transient knockdown of menin expression using a human MEN1 siRNA (Santa Cruz Biotechnology) (sc-35922) along with siRNA Transfection Reagent (Santa Cruz Biotechnology) (sc-29528), according to the vendor's instructions.

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Expressing, Flow Cytometry

Journal: The American Journal of Pathology

Article Title: miR-24 Inhibition Increases Menin Expression and Decreases Cholangiocarcinoma Proliferation

doi: 10.1016/j.ajpath.2016.10.021

Figure Lengend Snippet: Increased menin expression decreases proliferation. Mz-ChA-1 cells overexpressing menin with pCMV6-MEN1 vector exhibit a decrease in Ki-67 proliferative marker expression. A–C: Increased menin expression in pCMV6-MEN1 Mz-ChA-1 cells by real-time PCR (A) and flow cytometry (B) decreased Ki-67 proliferative marker expression by real-time PCR (C). D: Decreased cell migration as measured by wound healing assay. E: Decreased cell invasion as measured by Boyden chamber assay in pCMV6-MEN1 Mz-ChA-1 cells. Data are expressed as means ± SEM performed in triplicate (A–E). ∗P < 0.05 versus Mz-ChA-1 control cells. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Modulation of Menin Expression, Proliferation, and Angiogenesis in Vitro Mz-ChA-1 cells were targeted for the transient knockdown of menin expression using a human MEN1 siRNA (Santa Cruz Biotechnology) (sc-35922) along with siRNA Transfection Reagent (Santa Cruz Biotechnology) (sc-29528), according to the vendor's instructions.

Techniques: Expressing, Plasmid Preparation, Marker, Real-time Polymerase Chain Reaction, Flow Cytometry, Migration, Wound Healing Assay, Boyden Chamber Assay

Journal: The American Journal of Pathology

Article Title: miR-24 Inhibition Increases Menin Expression and Decreases Cholangiocarcinoma Proliferation

doi: 10.1016/j.ajpath.2016.10.021

Figure Lengend Snippet: Menin expression negatively regulates angiogenesis. A: By real-time PCR, Mz-ChA-1 MEN1 knockout cells increased expression of angiogenic factors compared to Mz-ChA-1 control cells. B: By real-time PCR, pCMV6-MEN1 Mz-ChA-1 cells decreased expression of angiogenic factors compared to Mz-ChA-1 control cells. Data are expressed as means ± SEM performed in triplicate (A and B). ∗P < 0.05 versus Mz-ChA-1 control cells. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Modulation of Menin Expression, Proliferation, and Angiogenesis in Vitro Mz-ChA-1 cells were targeted for the transient knockdown of menin expression using a human MEN1 siRNA (Santa Cruz Biotechnology) (sc-35922) along with siRNA Transfection Reagent (Santa Cruz Biotechnology) (sc-29528), according to the vendor's instructions.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Knock-Out

Journal: The American Journal of Pathology

Article Title: miR-24 Inhibition Increases Menin Expression and Decreases Cholangiocarcinoma Proliferation

doi: 10.1016/j.ajpath.2016.10.021

Figure Lengend Snippet: miR-24 negatively regulates menin. A: Real-time PCR evaluation of miR-24 expression in CCA and H69 cell lines demonstrates increased levels in CCA lines compared to H69 cells. B: Luciferase luminescence shows decreased menin expression with miR-24 mimic treatment. C: Left panel: miRNA-sequence data demonstrate increased expression of miR-24 in human CCA tumors compared with matched normal tissue. Right panel: Statistical significance of increased miR-24 expression is validated with an unpaired t-test. Data are expressed as means ± SEM (A–C). n = 3 (A and B); n = 9 (C). ∗P < 0.05 versus normal matched human control.

Article Snippet: Modulation of Menin Expression, Proliferation, and Angiogenesis in Vitro Mz-ChA-1 cells were targeted for the transient knockdown of menin expression using a human MEN1 siRNA (Santa Cruz Biotechnology) (sc-35922) along with siRNA Transfection Reagent (Santa Cruz Biotechnology) (sc-29528), according to the vendor's instructions.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Luciferase, Sequencing

Journal: The American Journal of Pathology

Article Title: miR-24 Inhibition Increases Menin Expression and Decreases Cholangiocarcinoma Proliferation

doi: 10.1016/j.ajpath.2016.10.021

Figure Lengend Snippet: miR-24 drives proliferation. A: Real-time PCR confirmed knockdown of miR-24 in Mz-ChA-1 cells by hairpin inhibitor. miR-24 knockdown increased menin expression via fluorescence-activated cell sorting (B) and decreased expression of angiogenic factors via real-time PCR (C). Data are expressed as means ± SEM performed in triplicate unless otherwise stated (A–C). ∗P < 0.05 versus Mz-ChA-1 control cells. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Modulation of Menin Expression, Proliferation, and Angiogenesis in Vitro Mz-ChA-1 cells were targeted for the transient knockdown of menin expression using a human MEN1 siRNA (Santa Cruz Biotechnology) (sc-35922) along with siRNA Transfection Reagent (Santa Cruz Biotechnology) (sc-29528), according to the vendor's instructions.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Fluorescence, FACS

Journal: Cancers

Article Title: Menin and Menin-Associated Proteins Coregulate Cancer Energy Metabolism

doi: 10.3390/cancers12092715

Figure Lengend Snippet: Identification of MEN1- modulated genes in breast cancer cells. ( A ) RT-qPCR of MEN1 in T47D or MCF-7 cells treated with vehicle or MEN1 shRNA lentivirus ( n = 3). ( B ) Quantitative Western immunoassays (WES) of menin expression in T47D or MCF-7 cells treated with vehicle or MEN1 shRNA lentivirus ( n = 3). ( C ) Venn diagrams of differentially expressed genes (fold change ≥1.5 or ≤0.66) in T47D or MCF-7 cells after sh MEN1 knockdown compared with vehicle controls ( n = 2). ( D ) Pathway annotation analysis of MEN1 -upregulated and MEN1 -downregulated genes in T47D or MCF-7 cells using DAVID including cancer hallmark pathways. ( E ) Schematic illustration of five major metabolic pathways. ( F ) Expression heat maps of oxidative phosphorylation (OXPHOS) and glycolytic genes in both MEN1 knockdown T47D and MCF-7 cells (fold changes relative to vehicle controls). ( G ) Bar charts of the expression levels of representative OXPHOS and glycolytic genes affected by MEN1 knockdown in T47D or MCF-7 cells using RT-qPCR. Data are presented as mean ± S.D. Unpaired two-tailed Student’s t -test was used for statistics. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The full-length human MEN1 cDNA from pBABE hygro MEN1 WT (Addgene plasmid #11024) was cloned into pRetroX-mycBioID-MCS at Not1 and Mlu I site by the Gibson reaction (New England Biolabs, Ipswich, MA, USA) to obtain pRetroX-mycBirA-MEN1.

Techniques: Quantitative RT-PCR, shRNA, Western Blot, Expressing, Knockdown, Phospho-proteomics, Two Tailed Test

Journal: Cancers

Article Title: Menin and Menin-Associated Proteins Coregulate Cancer Energy Metabolism

doi: 10.3390/cancers12092715

Figure Lengend Snippet: Identification of menin-associated proteins (MAPs) in breast cancer cells. ( A ) and ( B ) WES of BirA-Menin fusion proteins ( A ) and biotin-labeled proteins ( B ) in total lysates of BirA-MEN1 BioID engineered T47D or MCF-7 cells after incubating with or without doxycycline and biotin. ( C ) Schematic purification and proteomic identification of MAPs using LC–MS/MS. ( D ) Heatmap of the quantification of 35 MAPs commonly shared in T47D and MCF-7 cells. MAPs further verified by WES immunoassays were indicated by arrows. ( E ) Network analysis of 35 MAPs in MCF-7 cells. The distance between menin and MAPs represented the quantitative ratio of each MAP and menin. MAPs marked in blue were further assayed by WES. ( F ) Nuclear or cytoplasmic lysates of BirA-MEN1 BioID engineered T47D or MCF-7 cells after streptavidin beads pull-down were detected by WES with antibodies against menin, KMT2A, MED12, WAPL, GATA3, LaminA/C, or GAPDH. FL, full length; SP, spliced form.

Article Snippet: The full-length human MEN1 cDNA from pBABE hygro MEN1 WT (Addgene plasmid #11024) was cloned into pRetroX-mycBioID-MCS at Not1 and Mlu I site by the Gibson reaction (New England Biolabs, Ipswich, MA, USA) to obtain pRetroX-mycBirA-MEN1.

Techniques: Labeling, Purification, Liquid Chromatography with Mass Spectroscopy

Journal: Cancers

Article Title: Menin and Menin-Associated Proteins Coregulate Cancer Energy Metabolism

doi: 10.3390/cancers12092715

Figure Lengend Snippet: Expression correlation relationship of menin/MAPs genes and OXPHOS/glycolytic genes. ( A ) Workflow of the in silico correlation analysis of gene expression in The Cancer Genome Atlas (TCGA) breast cancer cohort. ( B ) Heatmaps of the expression correlation between MEN1 /selected 4 MAP genes and OXPHOS genes (upper) or glycolytic genes (lower) in normal (N) and tumor (T) samples. The genes are arranged from the highest to the lowest according to gene expression correlation coefficients of MEN1 –OXPHOS genes or MEN1 –glycolytic genes in breast tumors. ( C ) and ( D ) Scatter plots and linear regression analyses of MEN1/selected MAPs expression and mean expression of OXPHOS genes ( C ) or glycolytic genes ( D ) in normal and tumor samples. ( E ) Violin plots (lower panel) shows the average expressions of the genes of OXPHOS complexes I-V and glycolysis in the samples of each of the corresponding 4 groups are shown as violin plots. Based on the median values (where ≥median is “high” and

Article Snippet: The full-length human MEN1 cDNA from pBABE hygro MEN1 WT (Addgene plasmid #11024) was cloned into pRetroX-mycBioID-MCS at Not1 and Mlu I site by the Gibson reaction (New England Biolabs, Ipswich, MA, USA) to obtain pRetroX-mycBirA-MEN1.

Techniques: Expressing, In Silico, Gene Expression

Journal: Cancers

Article Title: Menin and Menin-Associated Proteins Coregulate Cancer Energy Metabolism

doi: 10.3390/cancers12092715

Figure Lengend Snippet: Bioenergetic dynamics are regulated by menin and MAPs in T47D and MCF-7 cells. ( A ) and ( B ) Glycolytic and OXPHOS ATP productions in T47D ( A ) or MCF-7 ( B ) cells infected with vehicle, sh MEN1 , sh KMT2A , sh MED12 , sh WAPL , or sh GATA3 lentivirus. Statistics represented the difference of glycolytic or OXPHOS ATP production between shRNA knockdown and vehicle controls. ( C ) and ( D ) Bar charts representing mitochondrial functions in the single knockdown of MEN1 , KMT2A , MED12 , WAPL , or GATA3 and their vehicle control in T47D ( C ) or MCF-7 ( D ) cells. ( E ) Schematic summary of mitochondrial dynamics affected by the knockdown of MEN1 or MAPs. ( F , G ) Bar charts represented the glycolytic functions in T47D ( F ) or MCF-7 ( G ) cells subject to gene knockdown by sh MEN1 , sh KMT2A , sh MED12 , sh WAPL , or sh GATA3 lentivirus. ( H ) Schematic summary of glycolytic functions affected by the knockdown of MEN1 or MAPs. Data are presented as mean ± S.D. ( n = 15–20 technical-replicate wells). Statistical significance was performed by an unpaired two-tailed Student’s t -test between treated groups and corresponding controls. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The full-length human MEN1 cDNA from pBABE hygro MEN1 WT (Addgene plasmid #11024) was cloned into pRetroX-mycBioID-MCS at Not1 and Mlu I site by the Gibson reaction (New England Biolabs, Ipswich, MA, USA) to obtain pRetroX-mycBirA-MEN1.

Techniques: Infection, shRNA, Knockdown, Control, Two Tailed Test

Journal: Cancers

Article Title: Menin and Menin-Associated Proteins Coregulate Cancer Energy Metabolism

doi: 10.3390/cancers12092715

Figure Lengend Snippet: Integrity of the menin–KMT2A complex is required for OXPHOS functions. ( A ) WES of T47D or MCF-7 cells treated with DMSO or 1 μM MI-503 for 3 days (left). Relative protein expression normalized to the average of LaminA/C in WES (right). FL, full length; SP, spliced form. ( B ) Nuclear lysates of T47D or MCF-7 cells treated with DMSO or 1 μM of MI-503 for 3 days were immunoprecipitated with the menin antibody or IgG, and assayed by WES (upper). Relative protein expression in WES (lower). The protein expression in DMSO treated input was normalized as 1. FL, full length; SP, spliced form. ( C ) Glycolytic or OXPHOS ATP production in T47D or MCF-7 cells treated with 1 μM MI-503 for 0, 1, 3, 6, and 72 h, or DMSO control for 72 h. ( D , E ) Bar charts of the Seahorse mitochondrial stress test ( D ) and glycolytic stress test ( E ) on T47D or MCF-7 cells treated with 1 μM MI-503 for 0, 1, 3, 6, and 72 h, or DMSO for 72 h. Data are presented as mean ± S.D. ( n = 10–15 technical-replicate wells). An unpaired two-tailed Student’s t -test was used to determine statistical significance for the difference between MI-503-treated groups and its controls. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The full-length human MEN1 cDNA from pBABE hygro MEN1 WT (Addgene plasmid #11024) was cloned into pRetroX-mycBioID-MCS at Not1 and Mlu I site by the Gibson reaction (New England Biolabs, Ipswich, MA, USA) to obtain pRetroX-mycBirA-MEN1.

Techniques: Expressing, Immunoprecipitation, Control, Two Tailed Test

Journal: Cancers

Article Title: Menin and Menin-Associated Proteins Coregulate Cancer Energy Metabolism

doi: 10.3390/cancers12092715

Figure Lengend Snippet: MEN1 and OXPHOS expression are increased in breast circulating tumor cells (CTCs). ( A ) t-SNE profile plots and cell clustering of 93 CTCs from 5 breast cancer patients based on the single cell RT-qPCR expression profiling of 11 OXPHOS genes ( NDUFA7 , NDUFA11 , NDUFA13 , NDUFB7 , NDUFS7 , NDUFS8 , NDUFV1 , SDHA , SDHB , SDHC , and SDHD ). ( B ) Violin plots of MEN1 or selected MAPs expression, mean expression of 7 glycolytic genes ( ALDOA , ALDOC , ENO1 , PFKL , PFKP , PGK1 , and TPI1 ) or mean expression of 11 OXPHOS genes (aforementioned) in the five cell clusters. Statistical significance among clusters was carried out using the Duncan multi-range test. ( C ) Mean expression of 7 glycolytic genes and 11 OXPHOS genes in these 93 breast CTCs or in the TCGA primary breast cancer cohort. ( D ) Glycolytic and OXPHOS ATP productions of T47D or MCF-7 cells after circulation ( n = 6–10 technical replicates). ( E , F ) Mitochondrial ( E ) and glycolytic ( F ) functions of T47D or MCF-7 cells after circulation ( n = 5–9 technical-replicate wells). Statistics represented the difference between no circulating control and each treatment. Data are presented as mean ± S.D. An unpaired two-tailed Student’s t -test was used for statistical significance determination. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The full-length human MEN1 cDNA from pBABE hygro MEN1 WT (Addgene plasmid #11024) was cloned into pRetroX-mycBioID-MCS at Not1 and Mlu I site by the Gibson reaction (New England Biolabs, Ipswich, MA, USA) to obtain pRetroX-mycBirA-MEN1.

Techniques: Expressing, Quantitative RT-PCR, Control, Two Tailed Test