Journal: Nature Immunology

Article Title: Lipid asymmetry disruption by XKR8 orchestrates neutrophil extracellular trap formation and inhibits fungal infection

doi: 10.1038/s41590-026-02456-z

Figure Lengend Snippet: a , b , Quantification of PMA-induced NET formation in WT and XKR8-deficient BMNs, treated with or without EGTA from the beginning (1 mM) ( n = 10) ( a ), or treated with EGTA (1 mM) at indicated time points after PMA stimulation ( n = 9) ( b ). c , Evaluation of membrane tension using Flipper-TR by FLIM imaging. Representative images of Flipper-TR and annexin V in PMA-induced NETs (left) and quantification of Flipper-TR lifetime ( τ ) in PS-exposed (annexin V + ) versus non-exposed (annexin V − ) cells (right) ( n = 50 in WT control, n = 57 in KO control, n = 59 in PMA-WT-annexin V + , n = 62 in PMA-WT-annexin V − , n = 53 in PMA-KO). Red indicates annexin V-AF594; green shows Flipper-TR. Scale bars, 5 μm. d , Schematic diagram showing calcium channels highly expressed on mouse neutrophils along with their respective inhibitors or agonists. e – i , Quantification of NET formation in WT and XKR8-deficient neutrophils treated with inhibitors and/or agonists listed in d . e , PMA-induced NET formation with inhibitors targeting TRPM2 (JNJ-28583113, 10 μM), TRPM4 (9-phenanthrol, 30 μM), TRPM7 (carvacrol, 100 μg ml −1 ), TRPV2 ( SKF96365 , 20 μM), TRPV4 (HC067047, 1 μM), Piezo1 (GsMTx-4, 10 μM), and TRPC3 (Pyr3, 5 μM) for 6 h ( n = 10). f , PMA-induced NETs in WT and XKR8-deficient neutrophils treated with the agonists targeting TRPM2 (bilirubin, 10 μM), TRPM7 (naltriben (NTB), 50 μM), TRPV2 (cannabidiol (CBD), 10 μM), TRPV2 (probenecid, 1 mM), TRPV4 (GSK1016790A, 100 nM), Piezo1 (Yoda1, 10 μM), and TRPC3 (GSK1702934A, 100 μM) for 6 h ( n = 10). g , PMA-induced NETs in WT and XKR8-deficient neutrophils with both inhibitors and agonists against TRPV2 ( SKF96365 and CBD), TRPV4 (HC067047 and GSK1016790A), Piezo1 (GsMTx-4 and Yoda1), TRPC3 (Pyr3 and GSK1702934A) for 6 h ( n = 9). h , i , As in f , quantification of LPS-induced NETs ( h ) or C. albicans -induced NETs ( i ) in WT and XKR8-deficient neutrophils treated with the agonists targeting TRPV2 (CBD, 10 μM), TRPV2 (probenecid, 1 mM), TRPV4 (GSK1016790A, 100 nM), Piezo1 (Yoda1, 10 μM) and TRPC3 (GSK1702934A, 100 μM) for 6 h ( n = 10). NETs were measured by the area of decondensed chromatin DNA ( a , b , e – i ). Each dot represents one individual field of view ( a , b , e – i ) or a single cell ( c ). Data are presented as mean ± s.e.m. from one representative experiment of three ( c , e – i ) and four ( a , b ) independent experiments. Statistical analysis was performed using a two-way ANOVA ( a , b , e – i ) and one-way ANOVA ( c ). P values are shown on the graphs.

Article Snippet: Additional inhibitors, including S62845 (HY-100741), emricasan (HY-10396), 2-APB (B6643), SKF96365 (HY-100001), HC067047 (HY-100208), Pyr3 (HY-108465), JNJ-28583113 (HY-149143), 9-phenanthrol (HY-108457), carvacrol (HY-N0711), GsMTx-4 (HY-P1410), probenecid (HY-B0545), GSK1016790A (HY-19608), GSK1702934A (HY-111098), bilirubin (HY-N0323), Yoda1 (HY-18723) and NTB (HY-101302) were obtained from MedChemExpress.

Techniques: Membrane, Imaging, Control, Single Cell

Journal: Nature Immunology

Article Title: Lipid asymmetry disruption by XKR8 orchestrates neutrophil extracellular trap formation and inhibits fungal infection

doi: 10.1038/s41590-026-02456-z

Figure Lengend Snippet: a , b , Quantification of NETs in WT and XKR8-deficient BMNs or HL-60 cells treated with PMA (100 nM) in the presence of inhibitors listed in Fig. . a , PMA-induced NETs formation with inhibitors targeting TRPM2 (glutathione, GSH, 10 μM; JNJ-28583113, 10 μM), TRPM4 (9-phenanthrol, 30 μM), TRPM7 (Carvacrol, 100 μg/mL), SOCE (2-APB, 25 μM; CM-4620, 10 μM), TRPV2 ( SKF96365 , 20 μM), TRPV4 (HC067047, 1 μM), Piezo1 (GsMTx-4, 10 μM), TRPC3 (Pyr3, 5 μM), TRPC6 (SAR7334, 1 μM) ( n = 8-9). b , As in a , Quantification of PMA-induced NETs in WT and XKR8-deficient dHL-60 cells with inhibitors HC067047 (1 μM), Pyr3 (5 μM), SKF96365 (20 μM), 9-phenanthrol (30 μM), JNJ-28583113 (10 μM), Carvacrol (100 μg/mL) ( n = 8). c , As in a , quantification of NETs in human primary neutrophils isolated from healthy donors (HD, n = 8) treated with PMA (10 nM) in the presence of mechanosensitive Ca 2+ channel inhibitors. d , Calcium influx was evaluated in WT and XKR8 KO BMNs by Indo-1. Indo-1 signals were recorded immediately after adding the agonists targeting TRPV2 (Cannabidiol, CBD, 5 μM), TRPC3 (GSK1702934A, 100 μM), TRPV4 (GSK1016790A, 100 nM), Piezo1 (Yoda1, 10 μM) for 5 min. A23187 (1.25 μM) as a positive control. e , Quantification of intracellular Ca 2+ levels in WT and XKR8 KO BMNs by Indo-1. Indo-1 fluorescence signals were measured at 2 h after treatment with PMA (100 nM) and in the presence of the agonists targeting TRPV2 (CBD, 5 μM), TRPC3 (GSK1702934A, 100 μM), TRPV4 (GSK1016790A, 100 nM), Piezo1 (Yoda1, 10 μM) ( n = 3). f , Quantification of PMA-induced NETs in WT and XKR8-deficient dHL-60 cells with agonists targeting TRPV2 (CBD, 5 μM; Probenecid, 1 mM), TRPC3 (GSK1702934A, 100 μM), TRPV4 (GSK1016790A, 100 nM), TRPM7 (Naltriben Mesylate, NTB, 50 μM) ( n = 8). g , h , Quantification of NETs in human primary neutrophils ( g , n = 8) or mouse BMNs ( h , n = 10) treated with agonists in the absence of PMA. i , Quantification of nigericin-induced NETs in WT and XKR8-deficient BMNs in the presence of the agonists targeting TRPV2 (CBD, 5 μM), TRPC3 (GSK1702934A, 100 μM), TRPV4 (GSK1016790A, 100 nM), Piezo1 (Yoda1, 10 μM) ( n = 10). j , k , Quantification of PMA-induced NETs ( j ) and PS exposure ( k ) in TRPV2, TRPC3, TRPV4, and Piezo1-deficient dHL-60 cells ( n = 10-12). NETs were quantified by a nuclear area greater than 80 µm 2 for BMNs ( a , c , g – i ) or 100 µm 2 for HL-60 cells ( b , f , j ). Each dot represents one individual field of view ( a – c , f – j ) or a biological replicate ( e , k ). Data are presented as mean ± SEM from one representative experiment of three ( a – k ) independent experiments. Statistical analysis was performed using two-way ANOVA ( a , b , f , h – k ), two-sided unpaired Student’s t-test ( e ), or one-way ANOVA ( c , g ). P values are shown on the graphs.

Article Snippet: Additional inhibitors, including S62845 (HY-100741), emricasan (HY-10396), 2-APB (B6643), SKF96365 (HY-100001), HC067047 (HY-100208), Pyr3 (HY-108465), JNJ-28583113 (HY-149143), 9-phenanthrol (HY-108457), carvacrol (HY-N0711), GsMTx-4 (HY-P1410), probenecid (HY-B0545), GSK1016790A (HY-19608), GSK1702934A (HY-111098), bilirubin (HY-N0323), Yoda1 (HY-18723) and NTB (HY-101302) were obtained from MedChemExpress.

Techniques: Isolation, Positive Control, Fluorescence

Journal: Cells

Article Title: Soluble β-Amyloid Oligomers Selectively Upregulate TRPC3 in Excitatory Neurons via Calcineurin-Coupled NFAT.

doi: 10.3390/cells14110843

Figure Lengend Snippet: Figure 1. TRPC3 expression is upregulated in human brains. (A,B) Representative Western blots of TRPC3 and TRPC6 protein expression from 9 AD and 7 non-AD control (NC) hippocampal tissue samples. ** p < 0.01. F: female; M: male. (C) Relative mRNA levels of Trpc3 and Trpc6 genes were determined by RT-qPCR based on 7 AD and 6 NC samples of human AD brain PFC regions; only RIN > 7 samples were used. * p < 0.05. (D) Representative microscopic images displaying immunostained amyloid pathology (with 4G8 antibody). (E) Representative TRPC3-immunostained microscopic images of AD and control brains. Red arrowheads indicate typical cells with morphologies of large pyramidal neurons, respectively. (F) Quantification of TRPC3 DAB immunosignals based on 5 AD and 4 NC cases. ** p < 0.01. Note: human brain samples used for panels (A-C) were from same cohort of patient and control individuals, and representative images shown in panels (D,E) were taken from same pair of AD and NC cases from different cohort of clinical samples.

Article Snippet: The TRPC3 agonist GSK1702934A (GSK) (cat. # 6508, Tocris, Ellisville, MI, USA) and fast voltage-gated sodium channel blocker tetrodotoxin citrate (TTX) (cat. # 1069, Tocris) were each bath-applied.

Techniques: Expressing, Western Blot, Control, Quantitative RT-PCR

Journal: Cells

Article Title: Soluble β-Amyloid Oligomers Selectively Upregulate TRPC3 in Excitatory Neurons via Calcineurin-Coupled NFAT.

doi: 10.3390/cells14110843

Figure Lengend Snippet: Figure 2. 7PA2 treatment upregulates TRPC3 and downregulates TRPC6 expression in cultured primary neurons (14 DIV). (A) Representative TRPC3 and TRPC6 immunostaining images of 7PA2- treated neurons. (B) Representative Western blots and quantification based on 3 independent experi- ments. * p < 0.05; ** p < 0.01. (C) Representative TRPC family immunoblots of cellular lysates from cultured neurons at 4 h after 7PA2 treatment.

Article Snippet: The TRPC3 agonist GSK1702934A (GSK) (cat. # 6508, Tocris, Ellisville, MI, USA) and fast voltage-gated sodium channel blocker tetrodotoxin citrate (TTX) (cat. # 1069, Tocris) were each bath-applied.

Techniques: Expressing, Cell Culture, Immunostaining, Western Blot

Journal: Cells

Article Title: Soluble β-Amyloid Oligomers Selectively Upregulate TRPC3 in Excitatory Neurons via Calcineurin-Coupled NFAT.

doi: 10.3390/cells14110843

Figure Lengend Snippet: Figure 3. 7PA2-induced TRPC3 upregulation is mediated by overactivated calcineurin (CaN). (A) Cal- cineurin’s phosphatase activity was induced by 2-fold, as determined 1 h after 7PA2 treatment. (n = 4). ** p < 0.01. (B) Representative TRPC3 immunocytochemistry showing preventive effects of two CaN inhibitors against 7PA2-upregulated TRPC3 expression (4 h time point). FK506 and CsA were used at 12.5 nM and 1 µM, respectively, in co-treatment with 7PA2. (C) Quantification of experiments of panel B (N = 3 independent experiments). * p < 0.05 and ** p < 0.01, respectively.

Article Snippet: The TRPC3 agonist GSK1702934A (GSK) (cat. # 6508, Tocris, Ellisville, MI, USA) and fast voltage-gated sodium channel blocker tetrodotoxin citrate (TTX) (cat. # 1069, Tocris) were each bath-applied.

Techniques: Activity Assay, Immunocytochemistry, Expressing

Journal: Cells

Article Title: Soluble β-Amyloid Oligomers Selectively Upregulate TRPC3 in Excitatory Neurons via Calcineurin-Coupled NFAT.

doi: 10.3390/cells14110843

Figure Lengend Snippet: Figure 4. 7PA2 treatment induces NFAT4c binding to Trpc3 gene promoter. (A,B) Schematic rep- resentation of 5′ upstream region of rat Trpc3 and Trpc6 genes. Sequences of rat Trpc3 gene (be- tween –1802 and –1217 bp) and rat Trip6 gene (between –812 and –771 bp) are aligned with cor- responding sequences of mouse and human genes. NFAT sites are shown in red. We identified two NFAT binding sites between positions –1802 and –1217 within Trpc3 promoter based on motif ([A/T]GGAAA[A/N][A/T/C]N) [57], as well as two NFAT binding sites from promoter Trpc6 [58]. These sites are relatively conserved among humans and rodents. Please note two differently spatially located sites within these two promoters. (C) 7PA2 treatment induces NFATc4 nuclear translocation as shown by immunocytochemistry. DAPI (4′,6-diamidino2-phenylindole, blue) staining is used to show colocalization of NFAT immunosignals at 2 h after 7PA2 treatment. (D) Representative (bottom) gel images of ChIP-PCR using NFATc4 antibody at 2h after 7PA2 treatment.

Article Snippet: The TRPC3 agonist GSK1702934A (GSK) (cat. # 6508, Tocris, Ellisville, MI, USA) and fast voltage-gated sodium channel blocker tetrodotoxin citrate (TTX) (cat. # 1069, Tocris) were each bath-applied.

Techniques: Binding Assay, Translocation Assay, Immunocytochemistry, Staining

Journal: Cells

Article Title: Soluble β-Amyloid Oligomers Selectively Upregulate TRPC3 in Excitatory Neurons via Calcineurin-Coupled NFAT.

doi: 10.3390/cells14110843

Figure Lengend Snippet: Figure 5. TRPC3 expression is upregulated in mature excitatory neurons upon 7PA2 treatment (4 h). (A,D) Representative immunocytochemical images of TRPC3 (A, green) and TRPC6 (D, green) co-stained with CaMKIIα (red) and DAPI (blue), taken at 4 h after 7PA2 or vehicle treatment. White arrowheads indicate excitatory neurons identified by high immunosignals of CaMKIIα, which are exclusively colocalized with strong TRPC3 immunosignals. (B) Quantification graphs based on number counts of cells expressing high and low levels of CaMKIIα co-localized with 7PA2- upregulated TRPC3. (C,E) These panels quantify cells of 7PA2-inuduced TRPC3 expression and 7PA2-reduced TRPC6 expression, respectively, in both high and low CaMKIIα populations (gray bars), compared to vehicle group (black bars). * p < 0.05; ** p < 0.01. (F) Representative fluorescent microscopic images of CaMKIIα and VGAT staining, revealing reciprocal staining patterns. White arrowheads indicate two neurons showing strong CaMKIIα immunosignals but weak VGAT, and yellow arrowheads indicate cells with opposite pattern.

Article Snippet: The TRPC3 agonist GSK1702934A (GSK) (cat. # 6508, Tocris, Ellisville, MI, USA) and fast voltage-gated sodium channel blocker tetrodotoxin citrate (TTX) (cat. # 1069, Tocris) were each bath-applied.

Techniques: Expressing, Staining

Journal: Cells

Article Title: Soluble β-Amyloid Oligomers Selectively Upregulate TRPC3 in Excitatory Neurons via Calcineurin-Coupled NFAT.

doi: 10.3390/cells14110843

Figure Lengend Snippet: Figure 6. Overactivated TRPC3 channel induces neuronal hyperexcitation in hippocampal CA1 region. (A) Representative microscopic image showing green fluorescent protein expression from the intra-CA1-injected AAV-eGFP viruses. (B) Western blot shows increased expression of TRPC3 from hippocampi-injected AAV-mTRPC3 and AAV-eGFP. Quantification was based on 3 independent experiments. * p < 0.05. (C) Increased TRPC3 expression and activation increase neuronal activities in CA1 region in brain slices (N = 5 mice). Left panels display example extracellular recordings of spontaneous neuronal activities in control side CA1 during baseline, during application of 5 µM GSK1702934A (GSK), and upon adding 1 µM tetrodotoxin (TTX). Data are quantified and summarized in bottom graph: each black dot is data point from one mouse, and red dots are the mean of black data points (5 mice, control side CA1). Right panels display same parameter measurements as left panel from AAV-mTRPC3-injected CA1 region. Scale bars apply to both panels. Before and after GSK frequency values in each sample for both groups were compared by paired t-test, whereas frequency values during GSK application in two groups were compared by unpaired t-test.

Article Snippet: The TRPC3 agonist GSK1702934A (GSK) (cat. # 6508, Tocris, Ellisville, MI, USA) and fast voltage-gated sodium channel blocker tetrodotoxin citrate (TTX) (cat. # 1069, Tocris) were each bath-applied.

Techniques: Expressing, Injection, Western Blot, Activation Assay, Control

Journal: Cells

Article Title: Soluble β-Amyloid Oligomers Selectively Upregulate TRPC3 in Excitatory Neurons via Calcineurin-Coupled NFAT.

doi: 10.3390/cells14110843

Figure Lengend Snippet: Figure 7. Selective blocking of TRPC3 alleviates 7PA2-induced synaptotoxicity. (A,B) Specificity of chosen siRNAs. (A) Quantified qRT-PCR results from neurons transfected with selected siRNAs against Trpc3 and Trpc6 genes. Neurons (5 DIV) were transfected with scrambled or specific siRNAs of Trpc3 and Trpc6 genes (sITRPC3 and siTRPC6). * p < 0.05 and ** p < 0.01. RT-qPCRs were performed in triplication using specific primers for detecting TRPC family genes. (B) Representative Western blot displays selectively downregulated expression of TRPC3 and TRPC6 proteins in transfected neurons (14 DIV). (* p < 0.05 and ** p < 0.01). (C) Representative MAP2 staining of neurons after being transfected with scrambled or specific siRNAs of Trpc3 and Trpc6 genes, without and with 7PA2 treatment. Quantification was based on 3 independent experiments. ** p < 0.01. (D) Co-treatment of neurons of 7PA2 with two TRPC3 selective antagonists (Pyr 3 and 10) significantly prevents dendritic loss. Quantification was based on 3 independent experiments. ** p < 0.01. (E) Co-treatment of neurons of 7PA2 with Pyr3 and 10 completely prevents synaptic Ca2+ flux induced by 7PA2. Quantification was based on over 20 neurons in each condition. Arrows indicate when compounds were perfused with and without 7PA2. “NS.” indicates non-significance and ** p < 0.01.

Article Snippet: The TRPC3 agonist GSK1702934A (GSK) (cat. # 6508, Tocris, Ellisville, MI, USA) and fast voltage-gated sodium channel blocker tetrodotoxin citrate (TTX) (cat. # 1069, Tocris) were each bath-applied.

Techniques: Blocking Assay, Quantitative RT-PCR, Transfection, Western Blot, Expressing, Staining

Journal: Journal of Cell Science

Article Title: Mammalian TRP ion channels are insensitive to membrane stretch

doi: 10.1242/jcs.238360

Figure Lengend Snippet: Purification of mTRPC6 ion channels for liposome reconstitution. (A) Size-exclusion chromatography profile of DDM-mTRPC6 protein complex corresponding to the tetramer state of the channel. AU, arbitrary unit. (B) SDS-PAGE showing stained mTRPC6 corresponding to the monomer state of the channel. The molecular mass standard is shown in kDa. (C) Illustration representing the patch excised from the liposome reconstituted with TRPC6.

Article Snippet: The chemical response test was performed at 21°C by placing a drop containing the TRPC6 agonist GSK1702934A (50 μM; Tocris Bioscience) or control buffer (M13 buffer: 30 mM Tris–HCl pH 7.0, 100 mM NaCl, 10 mM KCl, with 1% ethanol) in front of moving young adult hermaphrodites as described previously ( ; ).

Techniques: Purification, Size-exclusion Chromatography, SDS Page, Staining

Journal: Journal of Cell Science

Article Title: Mammalian TRP ion channels are insensitive to membrane stretch

doi: 10.1242/jcs.238360

Figure Lengend Snippet: Unidirectional incorporation of TRPC6 ion channel into the liposomal bilayer. (A) Representative hTRPC6 single-channel recording at different voltages in inside-out patches from HEK293T cells. Black arrowheads indicate the position of the baseline current. Upward and downward single-channel deflections represent outward and inward current, respectively. (B) Current-voltage relationship for single TRPC6 channels in inside-out patch mode ( n =6) from HEK293T cells. The single-channel conductance values are shown on the plot for inward (bottom left) and outward (top right) current: 44.6±4.7 pS and 24.1±3.1 pS respectively. (C) Amplitude histogram corresponding to the channel activity shown in A at +120 mV. (D) Open probability (NPo) of TRPC6 channel related to the patch voltages (mV) recorded in HEK293T cells. (E) Spontaneous activity of the Δ94 mTRPC6 channel in liposomes recorded at different voltages in the excised patch. Black arrowheads indicate the position of the baseline current. (F) Current voltage relationship of mTRPC6 recorded in liposomes with linear fitting ( n =5). Single-channel conductance values for inward and outward are 44.5±0.6 pS (bottom left) and 27.7±2.7 pS (top right), respectively. (G) Amplitude histogram corresponding to the channel activity shown in E, at −50 mV. (H) The relationship between voltage (mV) and open probability (NPo) of the TRPC6 recorded in liposomes. Other denotations are as in .

Article Snippet: The chemical response test was performed at 21°C by placing a drop containing the TRPC6 agonist GSK1702934A (50 μM; Tocris Bioscience) or control buffer (M13 buffer: 30 mM Tris–HCl pH 7.0, 100 mM NaCl, 10 mM KCl, with 1% ethanol) in front of moving young adult hermaphrodites as described previously ( ; ).

Techniques: Activity Assay, Liposomes

Journal: Journal of Cell Science

Article Title: Mammalian TRP ion channels are insensitive to membrane stretch

doi: 10.1242/jcs.238360

Figure Lengend Snippet: TRPC6 reconstituted in liposomes is insensitive to bilayer stretch. (A) Application of stretch to the liposome reconstituted with no protein control. (B) Representative trace of a constitutively active mTRPC6 channel that has been subjected to the negative pressure steps in the liposomes. The channel was recorded at indicated voltage in excised patch. The arrowhead indicates the closed state of the channel. (C) Application of the stretch to the MscL ion channel reconstituted into the liposomes, similar to the experiment shown in B. The downward deflections represent the inward current. The pressure values are shown under each trace. Black arrowheads indicate the position of the baseline current. (D) Summary of NPo change upon application of the stretch to the TRPC6 channel reconstituted in the liposomes ( n =6). For each applied pressure the NPo was calculated before and during pressure step. (E) Change of NPo upon application of negative pressure to MscL, similar to the experiment shown in D ( n =4). * P <0.05; ns, not significant, paired t -test. Other denotations are as in .

Article Snippet: The chemical response test was performed at 21°C by placing a drop containing the TRPC6 agonist GSK1702934A (50 μM; Tocris Bioscience) or control buffer (M13 buffer: 30 mM Tris–HCl pH 7.0, 100 mM NaCl, 10 mM KCl, with 1% ethanol) in front of moving young adult hermaphrodites as described previously ( ; ).

Techniques: Liposomes, Control

Journal: Journal of Cell Science

Article Title: Mammalian TRP ion channels are insensitive to membrane stretch

doi: 10.1242/jcs.238360

Figure Lengend Snippet: TRPC6 restores touch responses in osm-9 worms but requires diacylglycerol. (A) Schematic representation of the nose-touch response assay in worms. A C . elegans osm-9 mutant expressing Mus musculus (m)TRPC6 in sensory ASH neurons is subjected to eyelash stroke. (B) Schematic representation of chemical response test in a C . elegans osm-9 mutant expressing mTRPC6 in sensory ASH neurons. The freely moving worm is subjected to a drop of the mTRPC6 activator GSK1702934A (50 μM). The respective bar graphs of the right show the quantification (mean±s.e.m.) of the withdrawal responses upon eyelash stroke or application of GSK1702934A. n values for each condition are indicated within the bars. *** P <0.001; ns, not significant, Mann–Whitney test.

Article Snippet: The chemical response test was performed at 21°C by placing a drop containing the TRPC6 agonist GSK1702934A (50 μM; Tocris Bioscience) or control buffer (M13 buffer: 30 mM Tris–HCl pH 7.0, 100 mM NaCl, 10 mM KCl, with 1% ethanol) in front of moving young adult hermaphrodites as described previously ( ; ).

Techniques: Mutagenesis, Expressing, MANN-WHITNEY