Journal: Nature Communications

Article Title: Mitotic chromosome binding predicts transcription factor properties in interphase

doi: 10.1038/s41467-019-08417-5

Figure Lengend Snippet: Mitotic chromosome binding predicts genome-wide TF occupancy. a Correlation between the number of ChIP-seq peaks called and the ψon-rate for 8 different TFs. r -value (r) and p -value ( p ) are based on Pearson correlation. b Correlation between the number of ChIP-seq peaks called and TF relative residence times for 8 different TFs. r -value ( r ) and p -value ( p ) are based on Pearson correlation. c Genome tracks of RPKM-normalized bigWig files for each TF. The region encompasses the entire chromosome 19. The y -axis is scaled equally for all factors. d Correlation between the mitotic bound fraction and the number of ChIP-seq peaks called. Duplicates are indicated for OCT4 (yellow), SOX2 (green), BHLHB8 (red), and FOXA1 (dark blue). The two FOXA1 mutants are shown in light blue. Triangles TFs endogenously expressed in NIH-3T3. Circles: TFs not endogenously expressed in NIH-3T3. Rho-value (Rho) and p -value ( p ) are based on Spearman’s rank correlation. e Correlation between the mitotic bound fraction and the number of ChIP-seq peaks displaying the most frequently found motif for each TF, normalized over the total number of motif occurrences in the genome. Same color and shape coding as d . Rho-value (Rho) and p -value ( p ) are based on Spearman’s rank correlation. Error bars: SEM

Article Snippet: To compare the MBF between endogenous and exogenous expression of SOX2-SNAP, the cell lines were labeled with 24 nM of the SNAP-Cell_647-SiR dye (New England BioLabs Cat# S9102S) and imaged on the In Cell Analyzer with the Cy5 channel (Supplementary Fig. ).

Techniques: Binding Assay, Genome Wide, ChIP-sequencing

Journal: Nature Communications

Article Title: Mitotic chromosome binding predicts transcription factor properties in interphase

doi: 10.1038/s41467-019-08417-5

Figure Lengend Snippet: TF impact on chromatin accessibility as a function of occupancy and MBF. a Correlation between the mitotic bound fraction and the fraction of peaks in closed chromatin regions (devoid of ATAC-seq signal). Duplicates are indicated for OCT4 (yellow), SOX2 (green), BHLHB8 (red), and FOXA1 (dark blue). The two FOXA1 mutants are shown in light blue. Triangles: TFs endogenously expressed in NIH-3T3. Circles: TFs not endogenously expressed in NIH-3T3. r -value ( r ) and p -value ( p ) are based on Pearson correlation. b Example tracks of ChIP-seq signal, ATAC-seq signal in control cells, and ATAC-seq signal in TF overexpression cells for CDX2, FOXA1, and SIX6 for three regions with increased or decreased accessibility after TF overexpression (highlighted in red). The y-axis is scaled equally for ATAC-seq signal in control and TF overexpression within each region. c Correlation between the number of ChIP-seq peaks for each TF and the number of TF ChIP-seq peaks with significant change of chromatin accessibility upon overexpression of the TF. Known pioneers are indicated in green (SOX2) and blue (FOXA1). Rho-value (Rho) and p -value ( p ) are based on Spearman’s rank correlation. d Correlation between the mitotic bound fraction and the fraction of regions overlapping a ChIP-seq peak and displaying significant increase in their accessibility upon overexpression of the TF. Same color-coding as c . r -value ( r ) and p -value ( p ) are based on Pearson correlation. e Correlation between the mitotic bound fraction and the fraction of regions overlapping a ChIP-seq peak and displaying significant decrease in their accessibility upon overexpression of the TF. Same color-coding as c . Rho-value (Rho) and p -value ( p ) are based on Spearman’s rank correlation. Error bars: SEM

Article Snippet: To compare the MBF between endogenous and exogenous expression of SOX2-SNAP, the cell lines were labeled with 24 nM of the SNAP-Cell_647-SiR dye (New England BioLabs Cat# S9102S) and imaged on the In Cell Analyzer with the Cy5 channel (Supplementary Fig. ).

Techniques: ChIP-sequencing, Over Expression

Journal: Developmental biology

Article Title: Netrin1 establishes multiple boundaries for axon growth in the developing spinal cord

doi: 10.1016/j.ydbio.2017.08.001

Figure Lengend Snippet: (A–O) Thoracic level sections of E10.5 (A–E), E11.5 (F–J) or E12.5 (K–O) mouse spinal cords labeled with antibodies against Tag1 (red, A, B, F, G, K, L), Robo3 (red, D, E, I, J, N, O), neurofilament (NF, green) and Sox2 (blue).

Article Snippet: Immunohistochemistry The following primary antibodies were used overnight at 4°C: Rabbit: neurofilament (Cell Signaling Technology #C28E10, 1:200), Sox2, Goat: Sox2 (Santa Cruz Biotechnology #17320, 1:2000), human-Robo3 (R&D #AF3076, 1:200), β-galactosidase (1:2000), netrin1 (R&D #AF1109, 1:500); Mouse: neurofilament (DSHB #3A10, 1:100), Sox2 (Santa Cruz Biotechnology E4 #365823, 1:1000), Nkx2.2 (DSHB #74.5 A5-s, 1:100), mAb Tag1 (DSHB #4D7, 1:100) ( Dodd et al., 1988 ); Guinea Pig: Olig2 ( Novitch et al., 2001 ) (1:20,000).

Techniques: Labeling

Journal: Developmental biology

Article Title: Netrin1 establishes multiple boundaries for axon growth in the developing spinal cord

doi: 10.1016/j.ydbio.2017.08.001

Figure Lengend Snippet: (A–K) Thoracic (A, C–F, G, I–J), lumbar (B) or cervical (H, K) level transverse sections of E11.5 (A) or E12.5 (C–H) netrin1lacZ/+ and E11.5 (B) or E12.5 (I–K) netrin1+/+ mouse spinal cords labeled with antibodies against β-gal (red, A, C–H), netrin1 (red, B, I–K: panel B and I was processed without antigen retrieval), NF (green), Sox2 (blue, C) and Tag1 (blue, A–B, E).

Article Snippet: Immunohistochemistry The following primary antibodies were used overnight at 4°C: Rabbit: neurofilament (Cell Signaling Technology #C28E10, 1:200), Sox2, Goat: Sox2 (Santa Cruz Biotechnology #17320, 1:2000), human-Robo3 (R&D #AF3076, 1:200), β-galactosidase (1:2000), netrin1 (R&D #AF1109, 1:500); Mouse: neurofilament (DSHB #3A10, 1:100), Sox2 (Santa Cruz Biotechnology E4 #365823, 1:1000), Nkx2.2 (DSHB #74.5 A5-s, 1:100), mAb Tag1 (DSHB #4D7, 1:100) ( Dodd et al., 1988 ); Guinea Pig: Olig2 ( Novitch et al., 2001 ) (1:20,000).

Techniques: Labeling

Journal: Neural regeneration research

Article Title: Multiple factors to assist human-derived induced pluripotent stem cells to efficiently differentiate into midbrain dopaminergic neurons.

doi: 10.4103/1673-5374.378203

Figure Lengend Snippet: Figure 2 ｜ The modified dual SMAD pathway inhibition protocol yielded a low proportion of dopaminergic neurons. (A) Representative images of human induced pluripotent stem cells (hiPSC) with immunostaining of pluripotency markers, octamer-binding transcription factor 4 (OCT4) and stage-specific embryonic antigen-4 (SSEA4). Scale bars: 50 μm. (B) Representative images of neural precursor cells (NPC) induced from hiPSC with immunostaining of SRY-box transcription factor 2 (SOX2) and Nestin. Scale bars: 50 μm. (C) Representative images of hiPSC-derived neurons with immunostaining of microtubule- associated protein 2 (MAP2) (green) and human nuclei (HuNu) antibody (red). Scale bars: 50 μm. (D) Analysis of neuronal differentiation efficiency (number of cells expressing HuNu: n = 443 cells in dual SMAD pathway inhibition protocol). (E) Representative images of hiPSC-derived neurons with immunostaining of markers for different neuronal subtypes: tyrosine hydroxylase (TH), gamma-aminobutyric acid (GABA), vesicular glutamate transporter 1 (VGLUT1) (all in green), and MAP2 (red) grown on mouse astrocytes. Scale bars: 50 μm. (F) Differentiation ratio of dopaminergic neurons, GABAergic neurons, and glutamatergic neurons were assessed based on the proportion of TH, GABA, and VGLUT1 positive cells among MAP2 positive cells, respectively. At least three independent replicates were performed of each experiment. SMAD: Suppressor of mother against decapentaplegic.

Article Snippet: Primary antibodies included: octamer-binding transcription factor 4 (OCT4), a marker used for the identification of iPSC (rabbit, 1:100, BioVision, Milpitas, CA, USA, Cat# 6765-100, RRID: AB_2936816), stage-specific embryonic antigen-4 (SSEA4), a marker used for identification of iPSC (mouse, 1:100, Invitrogen, Rockford, AL, USA, Cat# 41-4000, RRID: AB_2533506), SRY-box transcription factor 2 (SOX2), a marker used for identification of NPC (mouse, 1:500, R&D Systems, Minneapolis, MN, USA, Cat# MAB2018, RRID: AB_358009), Nestin, a marker used for identification of NPC (rabbit, 1:1000, Millipore, Darmstadt, Germany, Cat# ABD69, RRID: AB_2744681), microtubule-associated protein 2 (MAP2), a marker of dendrites (rabbit, 1:1000, Millipore, Burlington, MA, USA, Cat# AB5622, RRID: AB_91939), MAP2 (mouse, 1:1000, Millipore, Cat# AMAb91375, RRID: AB_2716657), beta III tubulin (TUJ1), a neuronal marker (mouse, 1:1000, Sigma-Aldrich, Cat# T8660, RRID: AB_477590), anti-human nuclei (HuNu), a marker of nuclei in human cells (mouse, 1:500, Millipore, Cat# MAB1281, RRID: AB_94090), anti-vesicular glutamate transporter 1 (VGLUT1), a glutamatergic neuronal marker (mouse, 1:250, Millipore, Cat# MAB5502, RRID: AB_262185), gamma-aminobutyric acid (GABA), a GABAergic neuronal marker (rabbit, 1:1000, Sigma-Aldrich, Cat# A2052, RRID: AB_477652), tyrosine hydroxylase (TH), a dopaminergic neuronal marker (rabbit, 1:1000, Millipore, Cat# AB152, RRID: AB_390204), TH (chicken, 1:500, Millipore, Cat# AB9702, RRID: AB_570923), FOXA2, a midbrain neuronal marker (rabbit, 1:500, Cell Signaling Technology, Danvers, MA, USA, Cat# 8186, RRID: AB_10891055), and paired like homeodomain 3 (PITX3), another midbrain neuronal marker (rabbit, 1:500, Millipore, Cat# AB5722, RRID: AB_91997).

Techniques: Modification, Inhibition, Immunostaining, Binding Assay, Derivative Assay, Expressing

Journal: Neural regeneration research

Article Title: Multiple factors to assist human-derived induced pluripotent stem cells to efficiently differentiate into midbrain dopaminergic neurons.

doi: 10.4103/1673-5374.378203

Figure Lengend Snippet: Figure 4 ｜ The SHH + FGF8 differentiation protocol yielded a high proportion of dopaminergic neurons, with a small fraction of neurons that expressed FOXA2, a marker of midbrain dopaminergic neurons. (A) Representative images of neural precursor cells (NPC) induced from human induced pluripotent stem cells (hiPSC) with immunostaining of Forkhead box protein A2 (FOXA2) and SRY-box transcription factor 2 (SOX2). Scale bars: 50 μm. (B) Representative images of hiPSC-derived neurons in the SHH + FGF8 protocol with immunostaining of microtubule- associated protein 2 (MAP2) (green) and human nuclei (HuNu) antibody (red). Scale bars: 50 μm. (C) Comparison of the differentiation efficiency of neurons in the SHH + FGF8 protocol and modified dual SMAD pathway inhibition protocol (number of cells expressing HuNu: n = 607 cells in SHH+FGF8 protocol, n = 443 cells in dual SMAD pathway inhibition protocol). (D) Representative images of hiPSC-derived neurons in the SHH + FGF8 protocol with immunostaining of tyrosine hydroxylase (TH) (green) and MAP2 (red). Scale bars: 50 μm. (E) The proportion of dopaminergic neurons in the SHH + FGF8 protocol and modified dual SMAD pathway inhibition protocol was determined by calculating the percentage of TH-positive cells in MAP2-positive cells. (number of cells expressing MAP2: n = 581 cells in SHH + FGF8 protocol, n = 339 cells in dual SMAD pathway inhibition protocol). (F) The white box area in the image is enlarged and displayed on the right. NPC were infected with a DAT-Cherry lentivirus and then cultured on a glial cell feeder layer to label dopaminergic neurons in the later differentiation stage. Representative images of hiPSC-derived neurons in the SHH + FGF8 protocol with immunostaining of TH (green) and beta III tubulin (TUJ1) (white) on day 33. Scale bars: 50 μm (left), 10 μm (right). (G) The white box area in the image is enlarged and displayed at the bottom. Immunofluorescence staining of FOXA2 (red) and TH (green). Scale bars: 20 μm (upper panels), 5 μm (lower panels). At least three independent replicates were performed for each experiment. Data are presented as mean ± SEM, ****P < 0.0001. Statistical significance was evaluated by two-tailed unpaired Student’s t-test (C, E). FGF8: Fibroblast growth factor 8; ns: not significant; SHH: Sonic Hedgehog; TH: tyrosine hydroxylase.

Article Snippet: Primary antibodies included: octamer-binding transcription factor 4 (OCT4), a marker used for the identification of iPSC (rabbit, 1:100, BioVision, Milpitas, CA, USA, Cat# 6765-100, RRID: AB_2936816), stage-specific embryonic antigen-4 (SSEA4), a marker used for identification of iPSC (mouse, 1:100, Invitrogen, Rockford, AL, USA, Cat# 41-4000, RRID: AB_2533506), SRY-box transcription factor 2 (SOX2), a marker used for identification of NPC (mouse, 1:500, R&D Systems, Minneapolis, MN, USA, Cat# MAB2018, RRID: AB_358009), Nestin, a marker used for identification of NPC (rabbit, 1:1000, Millipore, Darmstadt, Germany, Cat# ABD69, RRID: AB_2744681), microtubule-associated protein 2 (MAP2), a marker of dendrites (rabbit, 1:1000, Millipore, Burlington, MA, USA, Cat# AB5622, RRID: AB_91939), MAP2 (mouse, 1:1000, Millipore, Cat# AMAb91375, RRID: AB_2716657), beta III tubulin (TUJ1), a neuronal marker (mouse, 1:1000, Sigma-Aldrich, Cat# T8660, RRID: AB_477590), anti-human nuclei (HuNu), a marker of nuclei in human cells (mouse, 1:500, Millipore, Cat# MAB1281, RRID: AB_94090), anti-vesicular glutamate transporter 1 (VGLUT1), a glutamatergic neuronal marker (mouse, 1:250, Millipore, Cat# MAB5502, RRID: AB_262185), gamma-aminobutyric acid (GABA), a GABAergic neuronal marker (rabbit, 1:1000, Sigma-Aldrich, Cat# A2052, RRID: AB_477652), tyrosine hydroxylase (TH), a dopaminergic neuronal marker (rabbit, 1:1000, Millipore, Cat# AB152, RRID: AB_390204), TH (chicken, 1:500, Millipore, Cat# AB9702, RRID: AB_570923), FOXA2, a midbrain neuronal marker (rabbit, 1:500, Cell Signaling Technology, Danvers, MA, USA, Cat# 8186, RRID: AB_10891055), and paired like homeodomain 3 (PITX3), another midbrain neuronal marker (rabbit, 1:500, Millipore, Cat# AB5722, RRID: AB_91997).

Techniques: Marker, Immunostaining, Derivative Assay, Comparison, Modification, Inhibition, Expressing, Infection, Cell Culture, Immunofluorescence, Staining, Two Tailed Test

Journal: Cell Death & Disease

Article Title: p97/VCP is highly expressed in the stem-like cells of breast cancer and controls cancer stemness partly through the unfolded protein response

doi: 10.1038/s41419-021-03555-5

Figure Lengend Snippet: a Immunoblotting analysis of SOX2 from the MDA-MB-231 cells treated with Eer I or NMS-873. GAPDH serves as a loading control. b Immunoblotting analysis of c-MYC from Eer I and NMS-873 treated MDA-MB-231 cells. GAPDH serves as a loading control. c Immunoblotting analysis of c-MYC from the MDA-MB-231 cells with p97 silenced. β-actin serves as a loading control. d c-MYC expression correlates with that of p97 in human breast cancer tissues. Left: representative immunohistochemical staining of c-MYC and p97 in the consecutive serial sections of breast cancer tissues. c-MYC and p97 expressions were classified as low and high. Bar: upper 200 μm, lower 50 μm. Right: correlation analyses of c-MYC and SOX2 expression in breast cancer tissues ( n = 98). e , f Immunoblotting analysis of SKP2 from Eer I and NMS-873 treated MDA-MB-231 cells or MDA-MB-231 cells with p97 silenced. GAPDH serves as a loading control. g SKP2 expression correlates with that of p97 in human breast cancer tissues. Left: representative immunohistochemical staining of SKP2 and p97 in the consecutive serial sections of breast cancer tissues. SKP2 and p97 expressions were classified as low or high. Bar: upper 200 μm, lower 50 μm. Right: correlation analyses of SKP2 and p97 expression in breast cancer tissues ( n = 75). h Immunoblotting analysis of SOX2 and SKP2 from mammospheres and adherent MDA-MB-231 cells. β-actin serves as a loading control. i Immunoblotting analysis of SOX2 and SKP2 from mammospheres with SKP2 silenced. β-actin serves as a loading control. j Mammosphere formation of MDA-MB-231 cells with SKP2 silenced. Left: the number of spheres under each treatment. Right: representative images. Bar: upper 100 μm. Data were shown as mean + SD. *** P < 0.001.

Article Snippet: The primary antibodies used in this study include: p97 (a generous gift from Dr. Hartmann Peterson, University of Copenhagen), ubiquitin (Santa Cruz, sc-8017), OCT4 (Proteintech, 11263-1-AP), SOX2 (Proteintech, 11064-1-AP), PERK (Cell Signaling Technology, 5683), phospho-PERK (Santa Cruz, sc-32577), ATF6α (Abcam, ab122897), BIP (Santa Cruz, sc-1051), XBP1 (BioLegend, 619502), ATF4 (Cell Signaling Technology, 11815), HIF-1α (Proteintech, 20960-1-AP), C/EBPδ (Santa Cruz, sc-636), c-MYC (Cell Signaling Technology, 5605), SKP2 (Proteintech, 11064-1-AP), β-actin (Molecular Biological Laboratories, PM053), and GAPDH (Cell Signaling Technology, 5174).

Techniques: Western Blot, Control, Expressing, Immunohistochemical staining, Staining

Journal: International Journal of Molecular Sciences

Article Title: Combined Dendritic and Axonal Deterioration Are Responsible for Motoneuronopathy in Patient-Derived Neuronal Cell Models of Chorea-Acanthocytosis

doi: 10.3390/ijms21051797

Figure Lengend Snippet: Characterization of the midbrain/hindbrain-differentiation protocol ( a ) Representative ICC images of ChAc patient iPSC derived mature midbrain/ hindbrain stem cell and neuronal cultures after three weeks. All cell lines show cells positive for stem cell/ early neuronal proteins (nestin, SOX2, TUBB3). Stainings for MAP2 and TH indicate the presence of mature dopaminergic neurons. Less than 1% of the cells express GALC or GFAP. ( b ) Quantification of IF images was done in Fiji, using macros which detected nuclei positive for each marker ( n ≥ 5). ( c ) Size comparison of MAP2 + and SMI32 + networks. Analysis was done in Fiji by skeletonizing thresholded IF images of MAP2/ SMI32 co-stainings and measuring the length of the obtained networks ( n ≥ 3). ( d ) Estimation of degeneration of the neuronal network. Fragmentation of MAP2 + and SMI32 + networks was calculated by assessing the network length and its perimeter and used as a surrogate for neurodegeneration ( n ≥ 3). ( e ) Decreased survival of ChAc patient iPSC derived mature midbrain/ hindbrain neurons after prolonged (6 months) cell culture. Cell amount was assessed by incubation of neuronal cultures grown in 96-wells with Hoechst 33342 and measuring the fluorescence using a Tecan Genios plate reader ( n ≥ 6). Scale bar = 100 µm, Boxes represent 25–75 percentiles, line represents median, whiskers represent 10–90%, + represents mean. Bars represent mean ± SEM. **/*** represents p < 0.01/0.001.

Article Snippet: In short, we transfected fibroblasts using four monocistronic retroviral vectors carrying the Yamanaka factors—OCT4, SOX2, KLF4 and MYCC (addgene #17217, #17218, #17219, #17220, #8449, #8454).

Techniques: Derivative Assay, Marker, Comparison, Cell Culture, Incubation, Fluorescence

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Transdifferentiating Astrocytes Into Neurons Using ASCL1 Functionalized With a Novel Intracellular Protein Delivery Technology

doi: 10.3389/fbioe.2018.00173

Figure Lengend Snippet: Small molecule only group after 2 days of priming by LDN193189 and SB431542 followed by 10 days of treatment with DAPT. (A) Cells express the early neural marker TUJ1 and weakly express the neural stem cell marker SOX2. Scale bar is 100 μm. (B) High magnification of (A) . Scale bar is 30 μm. (C) Cells are negative for the mature neural marker MAP2. Scale bar is 100 μm. (D) High magnification of (C) . Scale bar is 10 μm (E) Cells are negative for the mature neural marker NEUN and the neurotransmitter glutamate expressed by excitatory interneurons GLUT. Scale bar is 100 μm. (F) High magnification of (E) . Scale bar is 20 μm. (G) Cells are negative for the neurotransmitter tyrosine hydroxylase expressed by dopaminergic neurons TH, the neurotransmitter glutamic acid decarboxylase expressed by inhibitory interneurons GAD65/67, and the neurotransmitter choline acetyltransferase expressed by motor neurons CHAT. Scale bar is 100 μm. (H) High magnification of (G) . Scale bar is 30 μm. Note that some fluorescence is picked up by all the cells producing background fluorescence which can be ignored.

Article Snippet: The cells were stained for SOX2-PE Mouse IgG2A (R&D Systems) with IgG2A-PE isotype control (R&D Systems), TUJ1-PerCP Mouse IgG2A (R&D Systems) with IgG2A-PerCP isotype control (R&D Systems), MAP2-PE Mouse IgG1 (Clone AP20, Milli-Mark) with Mouse IgG1 PE isotype control (R&D Systems), and NEUN-PE Mouse IgG1 (Clone A60) with Mouse IgG1 PE isotype control (R&D Systems).

Techniques: Marker, Fluorescence

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Transdifferentiating Astrocytes Into Neurons Using ASCL1 Functionalized With a Novel Intracellular Protein Delivery Technology

doi: 10.3389/fbioe.2018.00173

Figure Lengend Snippet: Plastic astrocytes generate neurospheres and mature neurons after 12 days of exposure to ASCL1-IPTD, including 2 days of priming by LDN193189 and SB431542 followed by 10 days of DAPT. (A) Cells express the early neural marker TUJ1 and the neural stem cell marker SOX2. Scale bar is 100 μm. (B) High magnification of (A) . Scale bar is 30 μm. (C) Cells express the mature neural marker MAP2. Scale bar is 100 μm. (D) High magnification of (C) . Scale bar is 30 μm (E) Cells express the mature neural marker NEUN and the neurotransmitter glutamate expressed by excitatory interneurons GLUT. Scale bar is 100 μm. (F) High magnification of (E) . Scale bar is 30 μm. (G) Cells are negative for the neurotransmitter tyrosine hydroxylase expressed by dopaminergic neurons TH, and negative for the neurotransmitter choline acetyltransferase expressed by motor neurons CHAT, but do express the neurotransmitter glutamic acid decarboxylase expressed by inhibitory interneurons GAD65/67. Scale bar is 100 μm. (H) High magnification of (G) . Scale bar is 30μm. Note that some fluorescence is picked up by all the cells producing background fluorescence which can be ignored.

Article Snippet: The cells were stained for SOX2-PE Mouse IgG2A (R&D Systems) with IgG2A-PE isotype control (R&D Systems), TUJ1-PerCP Mouse IgG2A (R&D Systems) with IgG2A-PerCP isotype control (R&D Systems), MAP2-PE Mouse IgG1 (Clone AP20, Milli-Mark) with Mouse IgG1 PE isotype control (R&D Systems), and NEUN-PE Mouse IgG1 (Clone A60) with Mouse IgG1 PE isotype control (R&D Systems).

Techniques: Marker, Fluorescence