Journal: Journal of Biological Chemistry

Article Title: Inactivation of Max-interacting Protein 1 Induces Renal Cilia Disassembly through Reduction in Levels of Intraflagellar Transport 20 in Polycystic Kidney

doi: 10.1074/jbc.m112.413302

Figure Lengend Snippet: FIGURE 1. Identification of primary cilium in kidney of Mxi1-deficient mice. A, representative photographs of scanning electron micrographs of kidneys from wild-type (a–c), aged 9 months, and Mxi1-deficient kidney (d–f) as age-matched controls. B, comparison of the length of cilium in Mxi1-deficient kidney with wild-type kidney. The length of cilium was quantified by measuring from ciliary tip to basal body (basal body is bottom and protruding region of cilia) using the scale bar indicated in the figure. The graph shows mean S.D. (error bars) of three independent experiments. The one-tailed p value is 0.0001, considered extremely significant (***). C, representative photographs of anti-acetylated -tubulin-stained Mxi1-deficient kidney (b and c) and wild-type kidney (a)(anti-acetylated-tubulinstaining(green),DAPIstaining(blue),Cy,cyst.Originalmagnification,3000.Scalebars,10m.D,Westernblotofproteinsrelated to B-Raf/MEK/ERK pathway in Mxi1-deficient kidney and wild-type. -Actin was used as a loading control.

Article Snippet: Cells were transfected with Mxi1 and Ets-1 small interfering RNA (siRNA; Santa Cruz Biotechnology) using Lipofectamine RNAi MAX (Invitrogen), according to the manufacturer’s protocol.

Techniques: Comparison, One-tailed Test, Staining, Control

Journal: Journal of Biological Chemistry

Article Title: Inactivation of Max-interacting Protein 1 Induces Renal Cilia Disassembly through Reduction in Levels of Intraflagellar Transport 20 in Polycystic Kidney

doi: 10.1074/jbc.m112.413302

Figure Lengend Snippet: FIGURE 2. Effect of Mxi1 on ciliogenesis and p-ERK level in MEFs. A, representative photographs of anti-acetylated -tubulin-stained Mxi1/, Mxi1/ MEFs. White arrow indicates cilia stained with anti-acetylated -tubulin. Anti-acetylated -tubulin staining is shown in green, DAPI staining is blue. Original magnification, 400. Scale bar, 20 m. B, quantification of ciliated cells in Mxi1/, Mxi1/ MEFs. To measure ciliated cells, a ratio of the number of cilia to the numberofnucleiwascalculatedandmultipliedby100.ThegraphshowsmeanS.D.(errorbars)ofthreeindependentexperiments.Theone-tailedpvalueis 0.001, considered extremely significant (**). C, Western blot of p-ERK and total ERK in Mxi1/ and Mxi1/ MEFs. -Actin was used as a loading control.

Article Snippet: Cells were transfected with Mxi1 and Ets-1 small interfering RNA (siRNA; Santa Cruz Biotechnology) using Lipofectamine RNAi MAX (Invitrogen), according to the manufacturer’s protocol.

Techniques: Staining, Western Blot, Control

Journal: Journal of Biological Chemistry

Article Title: Inactivation of Max-interacting Protein 1 Induces Renal Cilia Disassembly through Reduction in Levels of Intraflagellar Transport 20 in Polycystic Kidney

doi: 10.1074/jbc.m112.413302

Figure Lengend Snippet: FIGURE 4. Transient overexpression of Mxi1 has an effect on ciliary phenotype of Mxi1/ MEFs. A, verification of Mxi1 and Ift20 mRNA expression in Mxi1/ and Mxi1 transiently overexpressed Mxi1/ MEFs. The graphs show mean S.D. (error bars) in triplicate. B, representative photographs of anti- acetylated -tubulin-stained Mxi1/ and Mxi1 transiently overexpressed Mxi1/ MEFs. White arrows indicates cilia stained with anti-acetylated -tubulin. Original magnification, 400. Scale bars, 20 m. C, quantification of ciliated cells in Mxi1/ and Mxi1 transiently overexpressed Mxi1/ MEFs. To measure ciliated cells, the ratio of the number of cilia to the number of nuclei was calculated and multiplied by 100. The graph shows mean S.D. of three independent experiments. The one-tailed p value is 0.007, considered significant (**). D, Western blot of p-ERK and total ERK in Mxi1/, Mxi1/, and Mxi1 transiently overexpressed Mxi1/ MEFs. -Actin was used as a loading control.

Article Snippet: Cells were transfected with Mxi1 and Ets-1 small interfering RNA (siRNA; Santa Cruz Biotechnology) using Lipofectamine RNAi MAX (Invitrogen), according to the manufacturer’s protocol.

Techniques: Over Expression, Expressing, Staining, One-tailed Test, Western Blot, Control

Journal: Journal of Biological Chemistry

Article Title: Inactivation of Max-interacting Protein 1 Induces Renal Cilia Disassembly through Reduction in Levels of Intraflagellar Transport 20 in Polycystic Kidney

doi: 10.1074/jbc.m112.413302

Figure Lengend Snippet: FIGURE3.InfluenceofMxi1inactivationonIFT-relatedgeneexpression.A,verificationofIft20,Ift88,andKif3mRNAexpressioninMxi1-deficientmice.The graphs show mean S.D. (error bars) of three independent experiments. *, p 0.05; **, p 0.01. B, verification of Ift20, Ift88, and Kif3 mRNA expression in Mxi1-deficient MEFs. The graphs show mean S.D. of three independent experiments. *, p 0.05; ***, p 0.005.

Article Snippet: Cells were transfected with Mxi1 and Ets-1 small interfering RNA (siRNA; Santa Cruz Biotechnology) using Lipofectamine RNAi MAX (Invitrogen), according to the manufacturer’s protocol.

Techniques: Expressing

Journal: Journal of Biological Chemistry

Article Title: Inactivation of Max-interacting Protein 1 Induces Renal Cilia Disassembly through Reduction in Levels of Intraflagellar Transport 20 in Polycystic Kidney

doi: 10.1074/jbc.m112.413302

Figure Lengend Snippet: FIGURE5.InfluenceofMxi1onIft20expressionanditspromoteractivity.A,verificationofMxi1andIft20mRNAexpressioninmIMCD-3cells(controls),Mxi1 transiently overexpressed mIMCD-3 cells, and Mxi1 siRNA-treated mIMCD-3 cells. The graphs show mean S.D. (error bars) in triplicate. B, Western blot of Ift20 and Mxi1 level in Mxi1 siRNA-treated mIMCD-3 cells and Mxi1 transiently overexpressed mIMCD-3 cells compared with that in their respective controls. -Actin was used as a loading control. C, deletion constructs of the Ift20 5-flanking region and luciferase activity in mIMCD-3 and Mxi1 transiently overexpressed mIMCD-3cells.TherepresentativegraphshowsmeanS.D.ofthreeindependentassays.***,p0.005.TherightpartoftheluciferaseassayshowsMxi1mRNA expression in Mxi1 transiently overexpressed mIMCD-3 cells compared with that in control used in this luciferase assay. The graph shows mean S.D. in triplicate.

Article Snippet: Cells were transfected with Mxi1 and Ets-1 small interfering RNA (siRNA; Santa Cruz Biotechnology) using Lipofectamine RNAi MAX (Invitrogen), according to the manufacturer’s protocol.

Techniques: Western Blot, Control, Construct, Luciferase, Activity Assay, Expressing

Journal: Journal of Biological Chemistry

Article Title: Inactivation of Max-interacting Protein 1 Induces Renal Cilia Disassembly through Reduction in Levels of Intraflagellar Transport 20 in Polycystic Kidney

doi: 10.1074/jbc.m112.413302

Figure Lengend Snippet: FIGURE 6. Effect of Mxi1 on ciliogenesis and p-ERK level in mIMCD-3 cells. A, representative photographs of anti-acetylated -tubulin-stained mIMCD-3 cellsandMxi1siRNA-treatedmIMCD-3cells.Whitearrowsindicateciliastainedwithanti-acetylated-tubulin.Originalmagnification,1200.Scalebars,10m. B, quantification of ciliated cells in control siRNA-treated and Mxi1 siRNA-treated mIMCD-3 cells. To measure ciliated cells, a ratio of the number of cilia to the number of nuclei was calculated and multiplied by 100. The graph shows mean S.D. (error bars) of three independent experiments. The one-tailed p value is 0.0263, which was considered significant (*). C, Mxi1, p-ERK, and total ERK protein levels in mIMCD-3 and Mxi1 siRNA-treated mIMCD-3 cells. -Actin was used as the loading control.

Article Snippet: Cells were transfected with Mxi1 and Ets-1 small interfering RNA (siRNA; Santa Cruz Biotechnology) using Lipofectamine RNAi MAX (Invitrogen), according to the manufacturer’s protocol.

Techniques: Staining, Control, One-tailed Test

Journal: Journal of Biological Chemistry

Article Title: Inactivation of Max-interacting Protein 1 Induces Renal Cilia Disassembly through Reduction in Levels of Intraflagellar Transport 20 in Polycystic Kidney

doi: 10.1074/jbc.m112.413302

Figure Lengend Snippet: FIGURE 7. Ets-1 is a mediator of the regulatory mechanism between Mxi1 and Ift20. A, screening of known transcription factors that bind to the Ift20 promoterusingTRANSFACdatabase.B,lanes2–4,gelshiftassay.Lane6,supershiftassay.Lane1,negativecontrolforthegelshiftassay.Lane5,negativecontrol for the supershift assay. N.E., nuclear extracts; Comp, competitor. C, luciferase assay with construct of 0.5-kb wt (wild-type) and 0.5-kb mt (mutant) containing mutant sequence for the Ets-1 binding site in mIMCD-3 and Ets-1 transiently overexpressed mIMCD-3. The representative graph shows mean S.D. (error bars) of three independent assays. ***, p 0.005. The right part of the luciferase assay graph indicates Ets-1 mRNA expression in mock (0.5-kb wt), Ets-1 transiently overexpressed (TF) 0.5-kb wt, and Ets-1 transiently overexpressed (TF) 0.5-kb mt transfected mIMCD-3 cells used in this luciferase assay. The graph shows mean S.D. in triplicate. D, Western blot of Ets-1 and Ift20 protein levels in mIMCD-3 and Ets-1 siRNA-treated mIMCD-3 cells. -Actin was used as a loading control. E, Mxi1 and Ets-1 protein levels in Mxi1 transiently overexpressed mIMCD-3 and mIMCD-3 cells. -Actin was used as a loading control. F, Ets-1 and Mxi1 protein levels in mIMCD-3 cells, Ets-1 transiently overexpressed mIMCD-3 cells, and Ets-1 siRNA-treated mIMCD-3 cells.

Article Snippet: Cells were transfected with Mxi1 and Ets-1 small interfering RNA (siRNA; Santa Cruz Biotechnology) using Lipofectamine RNAi MAX (Invitrogen), according to the manufacturer’s protocol.

Techniques: Luciferase, Construct, Mutagenesis, Sequencing, Binding Assay, Expressing, Transfection, Western Blot, Control

Journal: Journal of Biological Chemistry

Article Title: Inactivation of Max-interacting Protein 1 Induces Renal Cilia Disassembly through Reduction in Levels of Intraflagellar Transport 20 in Polycystic Kidney

doi: 10.1074/jbc.m112.413302

Figure Lengend Snippet: FIGURE8.ProposedmodelillustratinglinksbetweeninactivationofMxi1 and renal cyst formation induced by cilia disassembly. Inactivation of Mxi1 decreases the level of Ets-1, which binds to the Ift20 promoter. Down- regulation of Ets-1 reduces Ift20 expression and induces cilia disassembly in renal epithelial cells. This abnormal cilia phenotype increases the p-ERK level and causes renal cyst formation.

Article Snippet: Cells were transfected with Mxi1 and Ets-1 small interfering RNA (siRNA; Santa Cruz Biotechnology) using Lipofectamine RNAi MAX (Invitrogen), according to the manufacturer’s protocol.

Techniques: Expressing

Journal: Genome biology

Article Title: BORIS/CTCFL epigenetically reprograms clustered CTCF binding sites into alternative transcriptional start sites.

doi: 10.1186/s13059-024-03175-0

Figure Lengend Snippet: Fig. 7 Opening of chromatin by BORIS facilitates binding of other transcriptional factors. a Scatter plot displaying the enrichment of 190 TF motifs at CTCF/BORIS binding sites that reprogrammed into active promoters, compared to transcriptionally silent CTCF/BORIS sites in NIH3T3 + BORIS (clone#2) cells. b MAZ and MXI1 motifs are significantly enriched at CTCF/BORIS binding sites converted into active promoters. c Genome browser view illustrates the recruitment of TBP, HCFC1, MXI1, and MAZ proteins at the CTCF site within the Rbpjl promoter, activated by BORIS binding in NIH3T3 + BORIS (clone#2) cells. The activated promoter is highlighted by a red open box. d Left panel: Scatter plot of normalized read counts (log10) for HCFC1 occupancy at the combined set of HCFC1 binding sites (46,287) in NIH3T3 + BORIS (clone#2) cells compared to the same genomic sites in EV cells. Right panel: Heatmap of CTCF (red), BORIS (blue), and HCFC1 (brown) occupancy at the 19,519 HCFC1 sites from the left panel (connected by red arrow). e TF motifs enriched at HCFC1 peaks (60 bp around the summit of peak) in NIH3T3 + EV versus NIH3T3 + BORIS (clone#2) cells. f Heatmap of BORIS (blue), TBP (purple), HCFC1 (brown), MXI1 (orange), and MAZ (green) occupancy at the 5871 CTCF/BORIS binding sites converted into active promoters in NIH3T3 + BORIS (clone#2) cells compared to NIH3T3 + EV cells from Fig. 4h. g Summary of epigenetic reprogramming: BORIS binding recruits SRCAP, which replaces H2A histone with H2A.Z, leading to the opening of chromatin around CTCF sites. This, in turn, attracts other TFs to bind and stimulate transcription, resulting in the conversion of transcriptionally inert CTCF sites into active promoters

Article Snippet: Rabbit polyclonal antibody against SRCAP (Kerafast, ESL103), rabbit polyclonal against HCFC1 (Novus Biologicals, NB100-68209), goat affinity-purified polyclonal antibody against Mxi1 (R&D Systems, AF4185), rabbit monoclonal to TATA-binding protein TBP (Abcam, ab220788), rabbit polyclonal anti-PHF8 antibody (Bethyl Laboratories, A301-772A), rabbit polyclonal to MAZ (Abcam, ab85725), rabbit polyclonal antibody against the region of histone H3 containing the trimethylated lysine 27 (H3K27me3) (Diagenode, C15410195), rabbit polyclonal antibody against histone H3, trimethylated at lysine 36 (H3K36me3) (Diagenode, C15410058), rabbit polyclonal antibody against histone H2A.Z (Abcam, ab4174), rabbit polyclonal to Histone H3 (acetyl K27) (Abcam, ab4729), rabbit polyclonal to Histone H3 (tri methyl K4) (Abcam, ab8580), anti-RNA Polymerase II Antibody, CTD Antibody, clone 8WG16 (Millipore, 05–952-I-100UG).

Techniques: Binding Assay

Journal: BMC Genomics

Article Title: Global analysis of gene expression in NGF-deprived sympathetic neurons identifies molecular pathways associated with cell death

doi: 10.1186/1471-2164-12-551

Figure Lengend Snippet: Selection of genes regulated by NGF withdrawal identified in this study

Article Snippet: Pre-optimised Taqman ® gapdh (Rn99999916_s1), hprt1 (Mn00446968_m1), txnip (Rn01533885_g1), trib3 (Rn00595314_m1), ddit3 (Rn00492098_g1), ndrg1 (Rn01506130_m1), and mxi1 (Rn00565846_m1) primer/probe sets were supplied by Applied Biosystems.

Techniques: Selection, Ubiquitin Proteomics, Translocation Assay, Binding Assay, Derivative Assay

Journal: BMC Genomics

Article Title: Global analysis of gene expression in NGF-deprived sympathetic neurons identifies molecular pathways associated with cell death

doi: 10.1186/1471-2164-12-551

Figure Lengend Snippet: The proteins encoded by the 5 selected genes increase in level after NGF withdrawal . A , Immunoblotting analysis of Trib3, Ddit3, Txnip, Ndrg1 and Mxi1 protein levels using extracts prepared from sympathetic neurons cultured in the presence or absence of NGF ± CEP-11004 for 16 hours. For all 5 genes, protein levels were significantly increased at 16 hours after NGF withdrawal. Representative images are shown. c-Jun is shown as a positive control. ERK levels are shown as a loading control. The blots were scanned on a densitometer to quantitate the levels of each protein and were normalised to the ERK loading control. The ratio of the level of each protein in the presence of NGF (+NGF) to the level of each protein after NGF withdrawal (-NGF) or in the presence of CEP-11004 was then calculated. B , Analysis of Trib3, Ddit3, Txnip, Ndrg1 and Mxi1 protein levels and subcellular localisation by immunocytochemistry. Sympathetic neurons were treated as indicated and fixed at 0, 2, 4, 8, 12, or 16 hours after NGF withdrawal ± CEP-11004. The cells were then stained with the appropriate primary antibody followed by a FITC-conjugated secondary antibody, and Hoechst dye to label the nuclear DNA. Images were collected using the same exposure time for each timepoint and the coverslips in each gene group were analysed in parallel. The scale bar represents 20 μm.

Article Snippet: Pre-optimised Taqman ® gapdh (Rn99999916_s1), hprt1 (Mn00446968_m1), txnip (Rn01533885_g1), trib3 (Rn00595314_m1), ddit3 (Rn00492098_g1), ndrg1 (Rn01506130_m1), and mxi1 (Rn00565846_m1) primer/probe sets were supplied by Applied Biosystems.

Techniques: Western Blot, Cell Culture, Positive Control, Control, Immunocytochemistry, Staining

Journal: British Journal of Cancer

Article Title: Oesophageal adenocarcinoma is associated with a deregulation in the MYC/MAX/MAD network

doi: 10.1038/sj.bjc.6604398

Figure Lengend Snippet: mRNA expression of MYC/MAX/MAD network genes in Barrett's metaplasia and oesophageal adenocarcinoma. qRT–PCR was used to examine expression of genes encoding c-MYC, MAD1, MXI1, MXI1-0 and MAX in Barrett's metaplasia (BM n =25) and oesophageal adenocarcinoma (OAC n =37). Graphs represent mean fold change relative to matched normal gastric control (G, normalised to one) ±1 s.e.m. * denotes significant change relative to G, ° denotes significant difference between BM and OAC ( P <0.05).

Article Snippet: Western blotting was performed as described previously ( Brookes et al , 2006 ) with a mouse monoclonal antibody to c-MYC (1 μ g ml −1 , clone 9E10, Applied Biosystems) or a rabbit polyclonal antibody to MAD1 (1 μ g ml −1 , clone C-19, Autogen Bioclear, Calne, Wiltshire, UK) or MXI1 (1 μ g ml −1 , clone G-16, Autogen Bioclear).

Techniques: Expressing, Quantitative RT-PCR

Journal: British Journal of Cancer

Article Title: Oesophageal adenocarcinoma is associated with a deregulation in the MYC/MAX/MAD network

doi: 10.1038/sj.bjc.6604398

Figure Lengend Snippet: MYC/MAX/MAD network protein expression in Barrett's metaplasia and oesophageal adenocarcinoma. Expression of c-MYC, MAD1 and MXI1 protein was examined in Barrett's metaplasia (BM n =6) and oesophageal adenocarcinoma (OAC n =15) with matched normal gastric mucosa (G) by western blotting. Immunoreactive bands were assessed by semi-quantitative densitometry. Expression in BM and OAC is expressed relative to G (normalised to one); cytokeratin 19 (CK19) was employed for normalisation of protein loading. A representative western blot for each protein is shown alongside densitometry data representing mean expression change ±1 s.e.m. * denotes significant change relative to G, ° denotes significant change between BM and OAC.

Article Snippet: Western blotting was performed as described previously ( Brookes et al , 2006 ) with a mouse monoclonal antibody to c-MYC (1 μ g ml −1 , clone 9E10, Applied Biosystems) or a rabbit polyclonal antibody to MAD1 (1 μ g ml −1 , clone C-19, Autogen Bioclear, Calne, Wiltshire, UK) or MXI1 (1 μ g ml −1 , clone G-16, Autogen Bioclear).

Techniques: Expressing, Western Blot

Journal: British Journal of Cancer

Article Title: Oesophageal adenocarcinoma is associated with a deregulation in the MYC/MAX/MAD network

doi: 10.1038/sj.bjc.6604398

Figure Lengend Snippet: Immunolocalisation of MYC/MAX/MAD network proteins in Barrett's metaplasia and oesophageal adenocarcinoma. Paraffin sections of normal oesophagus, normal gastric fundus, Barrett's metaplasia and oesophageal adenocarcinoma were subjected to immunohistochemistry using antibodies designed against c-MYC, MAD1, MXI1 and MAX. Magnification × 40.

Article Snippet: Western blotting was performed as described previously ( Brookes et al , 2006 ) with a mouse monoclonal antibody to c-MYC (1 μ g ml −1 , clone 9E10, Applied Biosystems) or a rabbit polyclonal antibody to MAD1 (1 μ g ml −1 , clone C-19, Autogen Bioclear, Calne, Wiltshire, UK) or MXI1 (1 μ g ml −1 , clone G-16, Autogen Bioclear).

Techniques: Immunohistochemistry

Journal: British Journal of Cancer

Article Title: Oesophageal adenocarcinoma is associated with a deregulation in the MYC/MAX/MAD network

doi: 10.1038/sj.bjc.6604398

Figure Lengend Snippet: Semi-quantitative analysis of c-MYC network proteins in the oesophagus

Article Snippet: Western blotting was performed as described previously ( Brookes et al , 2006 ) with a mouse monoclonal antibody to c-MYC (1 μ g ml −1 , clone 9E10, Applied Biosystems) or a rabbit polyclonal antibody to MAD1 (1 μ g ml −1 , clone C-19, Autogen Bioclear, Calne, Wiltshire, UK) or MXI1 (1 μ g ml −1 , clone G-16, Autogen Bioclear).

Techniques:

Journal: British Journal of Cancer

Article Title: Oesophageal adenocarcinoma is associated with a deregulation in the MYC/MAX/MAD network

doi: 10.1038/sj.bjc.6604398

Figure Lengend Snippet: Confirmation of expression in transfected SEG1 cells. SEG1 cells were transfected with pcDNA3.1/Zeo-MYCER, pcDNA3-MAD1 or the corresponding empty vector (VO); in the case of MYCER the chimeric product was then activated by the addition of 4-OHT. ( A ) qRT–PCR was utilised to assess MYC ( ) or MXD1 ( ) mRNA expression. ( B ) Western blotting demonstrated the expression the chimeric protein in SEG1-MYCER or MAD1 in SEG1-MAD1. Densitometric scanning approximated the fold increase in expression; a representative blot is also shown. Values represent the mean of two experiments each performed in triplicate ±1 s.e.m. * denotes statistical significance ( P <0.05).

Article Snippet: Western blotting was performed as described previously ( Brookes et al , 2006 ) with a mouse monoclonal antibody to c-MYC (1 μ g ml −1 , clone 9E10, Applied Biosystems) or a rabbit polyclonal antibody to MAD1 (1 μ g ml −1 , clone C-19, Autogen Bioclear, Calne, Wiltshire, UK) or MXI1 (1 μ g ml −1 , clone G-16, Autogen Bioclear).

Techniques: Expressing, Transfection, Plasmid Preparation, Quantitative RT-PCR, Western Blot

Journal: British Journal of Cancer

Article Title: Oesophageal adenocarcinoma is associated with a deregulation in the MYC/MAX/MAD network

doi: 10.1038/sj.bjc.6604398

Figure Lengend Snippet: c-MYC network expression in SEG1 cells expressing exogenous MYC/MAX/MAD network proteins. ( A ) qRT–PCR was utilised to evaluate the expression of MXD1 , MXI1 , MXI-0 and MAX mRNA in SEG1 cells transiently overexpressing MYCER. Relative gene expression is expressed as a ratio of SEG1-MYCER not stimulated using 4OHT normalised to one. ( B ) Expression of MYC , MXI1 , MXI-0 and MAX mRNA was assessed in SEG1 cells transiently overexpressing MAD1. Relative gene expression is expressed as a ratio of mock transfected cells normalised to one. Data represent the mean of two independent experiments each performed in triplicate ±1 s.e.m. * denotes statistical significance ( P <0.05).

Article Snippet: Western blotting was performed as described previously ( Brookes et al , 2006 ) with a mouse monoclonal antibody to c-MYC (1 μ g ml −1 , clone 9E10, Applied Biosystems) or a rabbit polyclonal antibody to MAD1 (1 μ g ml −1 , clone C-19, Autogen Bioclear, Calne, Wiltshire, UK) or MXI1 (1 μ g ml −1 , clone G-16, Autogen Bioclear).

Techniques: Expressing, Quantitative RT-PCR, Transfection