Journal: eLife

Article Title: SynGAP isoforms differentially regulate synaptic plasticity and dendritic development

doi: 10.7554/eLife.56273

Figure Lengend Snippet: ( A ) Schematic of SYNGAP1 splicing at the C-terminus. SYNGAP1 is alternatively spliced within exons 18–20 to generate four unique C-terminal isoforms designated as α1, α2, β, and γ. ( B ) C-terminal amino-acid sequences of SynGAP isoforms encoding select protein domains. Coil-Coil domain (yellow) and PDZ ligand-binding domain (blue). Targeted epitopes of isoform-specific SynGAP antibodies (JH2469, JH7265, JH7206, and JH7366) are indicated as dotted lines. ( C ) Specificity of SynGAP isoform-specific antibodies. Immunoblots of SynGAP isoform expression in lysates prepared from HEK 293 T cells expressing individual GFP-tagged SynGAP isoforms and lysates prepared from brain tissue obtained from WT and Syngap1 +/- mice were shown. Quantification of relative SynGAP isoform levels with respect to total SynGAP expression measured from immunoblot were shown in . Two-way ANOVA followed by Tukey's post hoc test (Isoform F(4,30) = 1.900; p=0.13, Genotype F(1,30) = 451.2; p<0.001, Interaction F(4,30)=1.900; p=0.13, n = 4 each condition) was performed. Error bar indicates ± SEM. ( D ) Endogenous expression and distribution of SynGAP isoforms in various organs. Immunoblots of qualitative distribution of SynGAP isoforms in lysates prepared from various organ tissues of WT mice were shown. Asterisks indicate non-specific bands that are also detected in tissue from knockout mice. Two-way ANOVA followed by Tukey's post hoc test (Tissue F(5,144) = 1433; p<0.0001, Isoform F(7,144) = 229.3; p<0.0001, Interaction F(35,144) = 25.45; p<0.0001, n = 4 each condition) was performed. Heat map of immunoblots was displayed in . The amount of protein in the brain is standardized as 1.0. ( E ) Western blot of endogenous levels of individual SynGAP isoforms and other synaptic proteins in lysates prepared from several brain regions obtained from WT and Syngap1 +/- mice. (OB: Olfactory bulb, CC: Cerebral cortex, Hip: Hippocampus, ST: Striatum, Th: Thalamus, Mid: Midbrain, Ce: Cerebellum). Two-way ANOVA followed by Tukey's post hoc test (Brain regions F(7, 264)=1048; p<0.0001, Molecules F(10,264) = 8.0 x 10 −12 ; p>0.9999, Interaction F(70.264) = 59.06; p<0.0001, n = 4 each condition) was performed. Graph showing the mean values of each signal was displayed in . ( F–H ) Developmental expression profiles of individual SynGAP isoforms and related synaptic proteins. ( F ) Immunoblots of SynGAP isoform expression measured in forebrain tissue lysates prepared from WT and Syngap1 +/- mice at different developmental ages. ( G ) Quantification of immunoblots representing relative enrichments along developmental stage. The mean values of each signal were plotted in the graph. ( H ) Quantification of absolute SynGAP isoform abundance at P0 and P42 from C and G . Error bars indicate ± SEM. Two-way ANOVA followed by Tukey's post hoc test (Developmental stage F(10,330) = 397.4; p<0.0001, Molecule F(9,330) = 2.116; p=0.027, Interaction F(90,330) = 26.18; p<0.0001, n = 4 each condition) was performed. ( I ) mRNA expression of the β and non-β SYNGAP1 isoforms across age in human dorsolateral prefrontal cortex. The relative portion of RNAseq reads spanning the exon 17–18 junction supporting either isoform was plotted against human age (post-conception weeks and years) with a linear regression. ( J ) mRNA expression of the α1, α2, and γ SYNGAP1 isoforms across age. The relative portion of RNAseq reads spanning the exon 18–19 junction (γ) or 18–20 (α1, α2) junction supporting each isoform was plotted against human age. Reads per 80 million mapped (RP80M) of RNAseq data are shown in .

Article Snippet: GFP-SynGAP-α2 and GFP-SynGAP-γ also exhibited enhanced sedimentation in the presence of myc-PSD-95, albeit to a lesser extent than that of GFP-SynGAP-α1 (50.8 ± 0.9% of SynGAP-α2 and 53.3 ± 0.5% of SynGAP-γ in pellet fraction when co-expressed with PSD-95, ) These isoforms harbor a complete coiled-coil domain but lack the PDZ ligand.

Techniques: Ligand Binding Assay, Western Blot, Expressing, Knock-Out

Journal: eLife

Article Title: SynGAP isoforms differentially regulate synaptic plasticity and dendritic development

doi: 10.7554/eLife.56273

Figure Lengend Snippet: ( A ) (Left) Representative confocal time course images of dendritic spines of cultured neurons before and after photobleaching of shRNA-resistant GFP-SynGAP (WT or LDKD) under conditions of endogenous SynGAP knockdown. The left-most images display and overlay of the GFP signal from GFP-SynGAP and mCherry reporting the transfection of the SynGAP shRNA, two minutes before photobleaching. Time course images show relative fluorescence intensity before (t = −2 min), immediately following (t = 0 min), and 40 min (t = 40 min) following photobleaching with a 488 nm laser. Scale bar = 2 μm. (Right) Quantification of fluorescence recovery following photobleaching normalized to pre-bleach mean fluorescence intensity. Dashed-line curves (black = GFP SynGAP WT; red = GFP SynGAP LDKD) represent the results of single-order exponential fitting following nonlinear regression analysis. Shadows around each curve represent the SEM for the raw values. Curve plateaus were used to estimate the mobile fraction. The plateau of GFP-SynGAP WT recovery was 0.183 (95% CI = 0.169–0.199), and the plateau of GFP-SynGAP LDKD recovery was 0.372 (95% CI = 0.362–0.384). Plateaus were compared using the extra sum-of-squares F test. GFP-SynGAP WT n = 16 bleached spines from 4 neurons; GFP-SynGAP LDKD n = 19 bleached spines from 3 neurons. ***p<0.0001, F = 82.99 (1, 2026). ( B ) Representative immunoblot probing levels of GFP-SynGAP (WT or PDZ deletion mutant) and myc-PSD95 in phase-separated supernatant and pellet lysate fractions obtained from HEK cells expressing myc-PSD95 and either GFP-SynGAP-WT or GFP-SynGAP-ΔQTRV (PDZ ligand deletion) constructs. (Right panel) Quantification of pellet fraction ratios obtained from averaged immunoblots as the representative example shown in left panel. Error bars indicate ± SEM. Two-way ANOVA followed by Tukey's post hoc test (Molecules F(1,24) = 195.00; p<0.0001, Transfections F (3,24)=456.7; p<0.0001, Interaction F(3,24)=71.27; p<0.0001, n = 4, ***p<0.001, **p<0.01, *<0.05) was performed. ( C ) Representative confocal images of fixed HEK cells expressing myc-PSD95 alone or myc-PSD95 and either GFP-SynGAP-WT or GFP-SynGAP-LDKD constructs (F1). Scale Bar, 10 μm. Arrowheads indicate PSD95- and SynGAP-α1-containing puncta (>1 μm). (F2) Quantification of the averaged percentage of PSD95-positive puncta identified in images of fixed HEK cells as shown in (F1). Error bars indicate ± SEM. One-way ANOVA followed by Tukey's post hoc test (Transfections F(2, 9)=126.8; p<0.0001, n = 4 independent coverslip, ***p<0.001, **p<0.01, *<0.05) was performed.

Article Snippet: GFP-SynGAP-α2 and GFP-SynGAP-γ also exhibited enhanced sedimentation in the presence of myc-PSD-95, albeit to a lesser extent than that of GFP-SynGAP-α1 (50.8 ± 0.9% of SynGAP-α2 and 53.3 ± 0.5% of SynGAP-γ in pellet fraction when co-expressed with PSD-95, ) These isoforms harbor a complete coiled-coil domain but lack the PDZ ligand.

Techniques: Cell Culture, shRNA, Transfection, Fluorescence, Western Blot, Mutagenesis, Expressing, Construct

Journal: eLife

Article Title: SynGAP isoforms differentially regulate synaptic plasticity and dendritic development

doi: 10.7554/eLife.56273

Figure Lengend Snippet: ( A ) Efficiency of shRNA-SynGAP construct. Hippocampal neurons were transfected with shRNA-SynGAP#5 construct together with GFP (marker for transfected cell) and stained for pan-SynGAP antibody (**p<0.01, 77.3 ± 0.1% reduction of SynGAP expression upon our shRNA-SynGAP#5 construct, n = 4 cells, unpaired T-test (two-tailed), Error bars indicate ± SEM). Scale Bar, 10 μm. ( B ) Expression levels of our shRNA-resistant SynGAP isoform construct. Hippocampal neurons were transfected with mCherry and GFP-SynGAP isoform construct and stained for SynGAP isoform specific antibodies described in (α1 88.3 ± 0.2%, α2 97.0 ± 0.2%, β 83.5 ± 0.1% for GFP-SynGAP portion, n = 4 cells). Dotted line showed endogenous SynGAP expression levels. Error bars indicate ± SEM. One-way ANOVA followed by Tukey's post hoc test (Transfections F(4,10) = 126.8; p<0.01, n = 3 cells, *p<0.05) was performed. Scale Bar, 10 μm.

Article Snippet: GFP-SynGAP-α2 and GFP-SynGAP-γ also exhibited enhanced sedimentation in the presence of myc-PSD-95, albeit to a lesser extent than that of GFP-SynGAP-α1 (50.8 ± 0.9% of SynGAP-α2 and 53.3 ± 0.5% of SynGAP-γ in pellet fraction when co-expressed with PSD-95, ) These isoforms harbor a complete coiled-coil domain but lack the PDZ ligand.

Techniques: shRNA, Construct, Transfection, Marker, Staining, Expressing, Two Tailed Test

Journal: eLife

Article Title: SynGAP isoforms differentially regulate synaptic plasticity and dendritic development

doi: 10.7554/eLife.56273

Figure Lengend Snippet:

Article Snippet: GFP-SynGAP-α2 and GFP-SynGAP-γ also exhibited enhanced sedimentation in the presence of myc-PSD-95, albeit to a lesser extent than that of GFP-SynGAP-α1 (50.8 ± 0.9% of SynGAP-α2 and 53.3 ± 0.5% of SynGAP-γ in pellet fraction when co-expressed with PSD-95, ) These isoforms harbor a complete coiled-coil domain but lack the PDZ ligand.

Techniques: In Vitro, Recombinant, Sequencing, Clone Assay, shRNA, Bicinchoninic Acid Protein Assay, Software

Journal: The Journal of comparative neurology

Article Title: Distribution of the SynDIG4/ Proline rich transmembrane protein 1 in rat brain

doi: 10.1002/cne.23945

Figure Lengend Snippet: Primary antibodies Used in This Study

Article Snippet: Signal intensity in the blots co-probed with both antibodies is equivalent to the blots probed with single antibodies, indicating that the antibodies do not compete for the same binding site. table ft1 table-wrap mode="anchored" t5 caption a7 Target Host Isotype Type (clone) Immunogen Source Cat. No. RRID Form IB dilution ICC dilution IHC dilution SynDIG4 (L102/45) mouse IgG 2A mono (L102/45) Amino acids 1-66 rat MSSEKSGLPDSVPHTSPPPYNAPQPPAEPPIPPPQT APSSHHHHHHHYHQSGTATLPRLGAGGLAS Trimmer Lab (available at NeuroMab) 73-409 AB_2491106 TC supe Neat 1:2 1:2 SynDIG4 (NG5.1) rabbit poly RGPSSSATLPRPPH Smit Lab (GenScript) custom 1:1k 1:100 SynDIG4 (NG5.73) rabbit poly MSSEKSGLPDSVPH Smit Lab (GenScript) custom 1:1k 1:100 HA rat mono (3F10) Amino acids 76-111 of X47 hemaglutinin 1 YPYDVPDYA Roche 11867431001 AB_390919 purified 1:1k 1:100 β- tubulin mouse IgG 1 mono Amino acids 412-430 bovine SNMNDLVSEYQQYQDATA Millipore 05-661 AB_309885 protein G purified 1:5k SynDIG1 mouse IgG 2A mono (L42/17) Amino acids 1-183 of mouse MDGIIEQKSVLVHSKISDAGKRNGLINTRNFMAE SRDGLVSVYPAPQYQSHRLVASAAPGSLEGGRS EPVQQLLDPNTLQQSVESHYRPNIILYSDGVLRS WGDGVATDCCETTFIEDRSPTKDSLEYPDGKFID LSGDDIKIHTLSYDVEEEEELQELESDYSSDTESE DNFLMMPPRDHLG Trimmer Lab (available at NeuroMab) 75-251 AB_10999753 Purified IgG 1:1k 1:100 GluA1 rabbit poly human cytoplasmic domain Millipore AB1504 AB_2113602 1:3k 1:100 1:200 GluA2 mouse IgG 1 mono (L21/32) Amino acids 834-883 rat FCYKSRAEAKRMKVAKNAQNINPSSSQNSQNFA TYKEGYNVYGIESVKI NeuroMab 75-002 AB_2232661 Purified IgG 1:3k 1:100 PSD-95 mouse IgG 2A mono (K28/43) Amino acids 77-299 human PSD-95 FSIAGGTDNPHIGDDPSIFITKIIPGGAAAQDGRLR VNDSILFVNEVDVREVTHSAAVEALKEAGSIVRL YVMRRKPPAEKVMEIKLIKGPKGLGFSIAGGVG NQHIPGDNSIYVTKIIEGGAAHKDGRLQIGDKILA VNSVGLEDVMHEDAVAALKNTYDVVYLKVAK PSNAYLSDSYAPPDITTSYSQHLDNEISHSSYLGT DYPTAMTPTSPRRYSPVA NeuroMab 75-028 AB_2307331 Purified IgG 1:20k Synaptophysin 1 mouse IgG 1 mono (7.2) cytoplasmic tail Synaptic Systems 101011 AB_887824 Purified IgG 1:10k vGlut1 guinea pig poly C-terminus of rat vGlut1 Millipore AB5905 AB_2301751 serum 1:500 1:500 PSD-93 mouse IgG 1 mono (N18/30) full length rat NeuroMab 75-057 AB_2277296 Purified IgG 1:100 DAPI 1:2k Open in a separate window Primary antibodies Used in This Study All other primary antibodies used in this study are described in .

Techniques: Purification

Journal: The Journal of comparative neurology

Article Title: Distribution of the SynDIG4/ Proline rich transmembrane protein 1 in rat brain

doi: 10.1002/cne.23945

Figure Lengend Snippet: A: Representative blots of adult rat brains (six months old) subjected to sucrose gradient fractionation to enrich for the post-synaptic density (PSD). PSD-95 is used as a positive control to show enrichment at the PSD, and synaptophysin (Synapt) is used as a negative control to show absence in the PSD.

Article Snippet: Signal intensity in the blots co-probed with both antibodies is equivalent to the blots probed with single antibodies, indicating that the antibodies do not compete for the same binding site. table ft1 table-wrap mode="anchored" t5 caption a7 Target Host Isotype Type (clone) Immunogen Source Cat. No. RRID Form IB dilution ICC dilution IHC dilution SynDIG4 (L102/45) mouse IgG 2A mono (L102/45) Amino acids 1-66 rat MSSEKSGLPDSVPHTSPPPYNAPQPPAEPPIPPPQT APSSHHHHHHHYHQSGTATLPRLGAGGLAS Trimmer Lab (available at NeuroMab) 73-409 AB_2491106 TC supe Neat 1:2 1:2 SynDIG4 (NG5.1) rabbit poly RGPSSSATLPRPPH Smit Lab (GenScript) custom 1:1k 1:100 SynDIG4 (NG5.73) rabbit poly MSSEKSGLPDSVPH Smit Lab (GenScript) custom 1:1k 1:100 HA rat mono (3F10) Amino acids 76-111 of X47 hemaglutinin 1 YPYDVPDYA Roche 11867431001 AB_390919 purified 1:1k 1:100 β- tubulin mouse IgG 1 mono Amino acids 412-430 bovine SNMNDLVSEYQQYQDATA Millipore 05-661 AB_309885 protein G purified 1:5k SynDIG1 mouse IgG 2A mono (L42/17) Amino acids 1-183 of mouse MDGIIEQKSVLVHSKISDAGKRNGLINTRNFMAE SRDGLVSVYPAPQYQSHRLVASAAPGSLEGGRS EPVQQLLDPNTLQQSVESHYRPNIILYSDGVLRS WGDGVATDCCETTFIEDRSPTKDSLEYPDGKFID LSGDDIKIHTLSYDVEEEEELQELESDYSSDTESE DNFLMMPPRDHLG Trimmer Lab (available at NeuroMab) 75-251 AB_10999753 Purified IgG 1:1k 1:100 GluA1 rabbit poly human cytoplasmic domain Millipore AB1504 AB_2113602 1:3k 1:100 1:200 GluA2 mouse IgG 1 mono (L21/32) Amino acids 834-883 rat FCYKSRAEAKRMKVAKNAQNINPSSSQNSQNFA TYKEGYNVYGIESVKI NeuroMab 75-002 AB_2232661 Purified IgG 1:3k 1:100 PSD-95 mouse IgG 2A mono (K28/43) Amino acids 77-299 human PSD-95 FSIAGGTDNPHIGDDPSIFITKIIPGGAAAQDGRLR VNDSILFVNEVDVREVTHSAAVEALKEAGSIVRL YVMRRKPPAEKVMEIKLIKGPKGLGFSIAGGVG NQHIPGDNSIYVTKIIEGGAAHKDGRLQIGDKILA VNSVGLEDVMHEDAVAALKNTYDVVYLKVAK PSNAYLSDSYAPPDITTSYSQHLDNEISHSSYLGT DYPTAMTPTSPRRYSPVA NeuroMab 75-028 AB_2307331 Purified IgG 1:20k Synaptophysin 1 mouse IgG 1 mono (7.2) cytoplasmic tail Synaptic Systems 101011 AB_887824 Purified IgG 1:10k vGlut1 guinea pig poly C-terminus of rat vGlut1 Millipore AB5905 AB_2301751 serum 1:500 1:500 PSD-93 mouse IgG 1 mono (N18/30) full length rat NeuroMab 75-057 AB_2277296 Purified IgG 1:100 DAPI 1:2k Open in a separate window Primary antibodies Used in This Study All other primary antibodies used in this study are described in .

Techniques: Fractionation, Positive Control, Negative Control

Journal: eLife

Article Title: The palmitoyl acyltransferase ZDHHC14 controls Kv1-family potassium channel clustering at the axon initial segment

doi: 10.7554/eLife.56058

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-PSD93 (mouse monoclonal IgG1) , NeuroMab , Cat #75–057 (RRID: AB_2277296 ) , WB (1:500) IF (1:100).

Techniques: Recombinant, Construct, Transduction, shRNA, Sequencing, Plasmid Preparation, Transfection, Expressing, Isolation, Bicinchoninic Acid Protein Assay, Software

Journal: The Journal of Biological Chemistry

Article Title: Apolipoprotein E3 (ApoE3) but Not ApoE4 Protects against Synaptic Loss through Increased Expression of Protein Kinase C?

doi: 10.1074/jbc.M111.312710

Figure Lengend Snippet: ASPD-induced synaptic loss. A, confocal images of rat hippocampal primary neurons. Cells grown on chambered slides were treated with vehicle (Control), Aβ monomer (1 μm), ADDLs (1 μm), and 50 nm ASPD. Following a 20-h incubation, cells were stained for PSD-95 and synaptophysin. The first column represents the nucleus stained with DAPI (blue), the second column represents PSD-95 (green), the third column shows synaptophysin (red), and the fourth column is the merged image. Mean fluorescence intensity is expressed as a percentage of control (n = 6). Shown is a graphical representation of the expression level of PSD-95 (B) and synaptophysin (C). Values are mean ± S.E. (error bars) (Student's t test). *, p < 0.05; **, p < 0.005; ***, p < 0.0005.

Article Snippet: Primary antibodies (PKC-ϵ, β-actin, RACK1, synaptophysin, MAP-2, and PSD-95) were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

Techniques: Incubation, Staining, Fluorescence, Expressing

Journal: The Journal of Biological Chemistry

Article Title: Apolipoprotein E3 (ApoE3) but Not ApoE4 Protects against Synaptic Loss through Increased Expression of Protein Kinase C?

doi: 10.1074/jbc.M111.312710

Figure Lengend Snippet: ApoE3 prevents synaptic damage. Cells grown on chambered slides were treated with vehicle (Control), 50 nm ASPD, 50 nm ASPD + apoE3 (10 nm), 50 nm ASPD + apoE4 (10 nm), 50 nm ASPD + cholesterol (100 μm), 50 nm ASPD + apoE3 (10 nm) + cholesterol (100 μm), or ASPD (50 nm) + apoE4 (10 nm) + cholesterol (100 μm). Following a 20-h incubation, the cells were stained for MAP-2 (A) and synaptophysin (B) as described under “Experimental Procedures.” Mean fluorescence intensity is expressed as a percentage of control (n = 6). ApoE3 + cholesterol prevented the synaptic loss caused by ASPD. *, significance with respect to ASPD-treated cells. Error bars, S.E. *, p < 0.05.

Article Snippet: Primary antibodies (PKC-ϵ, β-actin, RACK1, synaptophysin, MAP-2, and PSD-95) were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

Techniques: Incubation, Staining, Fluorescence

Journal: The Journal of Biological Chemistry

Article Title: Apolipoprotein E3 (ApoE3) but Not ApoE4 Protects against Synaptic Loss through Increased Expression of Protein Kinase C?

doi: 10.1074/jbc.M111.312710

Figure Lengend Snippet: DCPLA-ME protects against ASPD-induced synaptic loss. A, rat hippocampal primary neurons grown on chambered slides were treated with vehicle (Control), 50 nm ASPD, 50 nm ASPD + 100 nm DCPLA-ME, and 50 nm ASPD + 100 nm DCPLA-ME + 5 μm PKCϵ translocation inhibitor. PKCϵ inhibitor was added 30 min before adding ASPD and DCPLA-ME. Following a 20-h incubation, cells were stained for MAP-2, PSD-95, and synaptophysin as described under “Experimental Procedures.” Mean fluorescence intensity was calculated and was expressed as a percentage of control (n = 6). ASPD treatment produced a marked decrease in stained neurite processes, whereas DCPLA-ME protected against synaptic loss. B, Western blot analysis of synaptophysin expression in control and ASPD-, ASPD + DCPLA-ME-, and PKCϵ inhibitor + ASPD + DCPLA-ME-treated primary rat hippocampal neurons. Values are mean ± S.E. (error bars) (Student's t test). *, p < 0.05; **, p < 0.005; ***, p < 0.0005.

Article Snippet: Primary antibodies (PKC-ϵ, β-actin, RACK1, synaptophysin, MAP-2, and PSD-95) were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

Techniques: Translocation Assay, Incubation, Staining, Fluorescence, Produced, Western Blot, Expressing

Journal: The Journal of Biological Chemistry

Article Title: Apolipoprotein E3 (ApoE3) but Not ApoE4 Protects against Synaptic Loss through Increased Expression of Protein Kinase C?

doi: 10.1074/jbc.M111.312710

Figure Lengend Snippet: ASPD specifically down-regulates PKCϵ in primary rat hippocampal neurons. A, PKCϵ, PKCα, and PKCδ mRNA were quantified by quantitative RT-PCR in control and ASPD-, apoE3 + cholesterol-, and ASPD + apoE3 + cholesterol-treated cells. Individual cDNA was amplified for PKCϵ and β-actin, and the PKCϵ signal was normalized to β-actin. PKCα and PKCδ showed no significant change on ASPD or apoE3 + cholesterol treatment. ApoE3 blocked the down-regulation by ASPD. B, RT-PCR shows that apoE4 has no effect on PKCϵ mRNA levels. C, RT-PCR shows that DCPLA-ME protects PKCϵ mRNA levels. D and E, immunoblot analysis of primary neurons treated with apoE3 (10 nm) + cholesterol (100 μm), DCPLA-ME (100 nm), PKCϵ inhibitor (5 μm), or 50 nm ASPD. Data are mean ± S.E. (error bars) of three independent experiments. *, significance with respect to control; #, significance with respect to ASPD-treated cells (Student's t test). *, p < 0.05; **, p < 0.005; ***, p < 0.0005.

Article Snippet: Primary antibodies (PKC-ϵ, β-actin, RACK1, synaptophysin, MAP-2, and PSD-95) were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

Techniques: Quantitative RT-PCR, Amplification, Reverse Transcription Polymerase Chain Reaction, Western Blot