rabbit anti phospho ship2  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Role of SHIP2 in cell repulsion regulated by Eph receptor and ephrin signaling

doi: 10.1101/872846

Figure Lengend Snippet: Experiments were carried out to analyse phosphorylation of SHIP2 at Y1135 after activation of EphB2 and/or ephrinB1. (A): EphB2 cells were incubated with soluble ephrinB1-Fc or ephrinB1 cells were incubated with soluble EphB2-Fc for 0, 5 and 15 min. The cells were lysed and phosphorylation of SHIP2 at Y1135 was measured by western blotting. Phosphorylation occurs after forward but not reverse signaling. (B): To understand the temporal relationship between EphB2 activation and SHIP2 Y1135 phosphorylation, a cell mixing assay was used. EphB2 or kiEphB2 cells were mixed with ephrinB1 cells, briefly centrifuged to force the cells into contact and incubated for 5, 15 or 30 min. EphB2 activation (phosphorylation on Y604) and phosphorylation of SHIP2 on Y1135 have a similar time course, and were not observed when a kinase inactive mutant of EphB2 was used.

Article Snippet: Antibodies used for Western blot analysis were to detect: EphB2 and GAPDH from Thermo Fisher (371700, MA5-15738, respectively); phosphorylated EphB2 from Abcam (ab61791); ephrinB1/2/3 from Santa Cruz (sc-910); FLAG tag from Sigma Aldrich (F3165); SHIP2 and phosphorylated SHIP2-Y1135 from Cell Signaling Technologies (#2839 and 5445, respectively).

Techniques: Activation Assay, Incubation, Western Blot, Mutagenesis

Journal: bioRxiv

Article Title: Role of SHIP2 in cell repulsion regulated by Eph receptor and ephrin signaling

doi: 10.1101/872846

Figure Lengend Snippet: Experiments were carried out to analyse the interaction between EphB2 and SHIP2. (A): HEK293 cells were transiently transfected with N-terminally tagged Flag-EphB2 construct. Flag-EphB2 was immunoprecipitated from the lysates and the precipitates analysed by western blotting. SHIP2 was found to co-immunoprecipitate with Flag-EphB2. (B): To determine if the interaction was driven by the SAM domain of SHIP2, Flag-SHIP2 (full length) and Flag-SHIP2 1-847 (delta SAM) was transiently transfected into EphB2 cells. Flag-SHIP2 (full length and delta SAM) was immunoprecipitated from the lysates. EphB2 was found to co-immunoprecipitate with both full length and delta SAM SHIP2. (C): To determine if the interaction is dependent on SH2 domain interactions, Flag-EphB2 and GFP-SHIP2 (full length) or GFP-SHIP2 1-120 were transiently co-transfected in HEK293 cells. Flag-EphB2 was immunoprecipitated from the lysates. The SH2 domain of SHIP2 is sufficient for the interaction with EphB2. (D): To test if the interaction is strengthened by EphB2 activation, Flag-SHIP2 was transiently expressed in EphB2 cells. The EphB2 cells were then mixed with either HEK293 cells (not activated) or mixed with ephrinB1 cells (activated). The cells were briefly centrifuged to force the cells into contact and incubated for 5 min before lysis. Flag-SHIP2 was immunoprecipitated from the lysates. Activation of EphB2 increased the interaction with SHIP2.

Article Snippet: Antibodies used for Western blot analysis were to detect: EphB2 and GAPDH from Thermo Fisher (371700, MA5-15738, respectively); phosphorylated EphB2 from Abcam (ab61791); ephrinB1/2/3 from Santa Cruz (sc-910); FLAG tag from Sigma Aldrich (F3165); SHIP2 and phosphorylated SHIP2-Y1135 from Cell Signaling Technologies (#2839 and 5445, respectively).

Techniques: Transfection, Construct, Immunoprecipitation, Western Blot, Activation Assay, Incubation, Lysis

Journal: bioRxiv

Article Title: Role of SHIP2 in cell repulsion regulated by Eph receptor and ephrin signaling

doi: 10.1101/872846

Figure Lengend Snippet: (A-H): Cell segregation assays were carried out in which EphB2 and ephrinB1 cells are mixed and plated at sub-confluent density on tissue culture dishes, then incubated for 48 h. EphB2 cells (green) and ephrinB1 cells (magenta) were labelled with cell tracker dyes, and colours changed using Fiji. (A) EphB2 / ephrinB1 control; (B) SHIP2 knockdown in EphB2 cells; (C) SHIP2 knockdown in ephrinB1 cells; (D) SHIP2 knockdown in both cell populations. (E-H): same assays as shown in the corresponding panel above, but with DAPI staining to visualise the compaction that occurs in EphB2 cell clusters. (I): quantitation of average EphB2 cell cluster size determined by thresholding and particle analysis in Fiji, normalised to control EphB2 / ephrinB1 assay. (J) Quantitation of border shift in boundary assays. Snapshots are shown in . Scale bars: 100 μm.

Article Snippet: Antibodies used for Western blot analysis were to detect: EphB2 and GAPDH from Thermo Fisher (371700, MA5-15738, respectively); phosphorylated EphB2 from Abcam (ab61791); ephrinB1/2/3 from Santa Cruz (sc-910); FLAG tag from Sigma Aldrich (F3165); SHIP2 and phosphorylated SHIP2-Y1135 from Cell Signaling Technologies (#2839 and 5445, respectively).

Techniques: Incubation, Staining, Quantitation Assay

Journal: bioRxiv

Article Title: Role of SHIP2 in cell repulsion regulated by Eph receptor and ephrin signaling

doi: 10.1101/872846

Figure Lengend Snippet: (A-D): boundary assays were carried out in which EphB2 and ephrinB1 cells meet to form a border. Snapshots are shown of the meeting of the two cell populations (A-D) and at the endpoint of the experiment (A’-D’; original position of border indicated with dashed white line). In control EphB2 / ephrinB1 assays (A, A’) the border shifts away from ephrinB1 cells. The border shift is not affected by knockdown of SHIP2 in EphB2 cells (B, B’) but decreases following knockdown in ephrinB1 cells (C, C’) or both populations (D, D’). The border shift is quantitated in . Scale bar: 100 μm.

Article Snippet: Antibodies used for Western blot analysis were to detect: EphB2 and GAPDH from Thermo Fisher (371700, MA5-15738, respectively); phosphorylated EphB2 from Abcam (ab61791); ephrinB1/2/3 from Santa Cruz (sc-910); FLAG tag from Sigma Aldrich (F3165); SHIP2 and phosphorylated SHIP2-Y1135 from Cell Signaling Technologies (#2839 and 5445, respectively).

Techniques:

Journal: bioRxiv

Article Title: Role of SHIP2 in cell repulsion regulated by Eph receptor and ephrin signaling

doi: 10.1101/872846

Figure Lengend Snippet: (A-C): Snapshots of time lapse movies (Movies 1-3, respectively) to illustrate cell repulsion response following interaction of EphB2 (green) and ephrinB1 (magenta) cells, with or without SHIP2 knockdown, indicated with arrowheads. Cell repulsion is characterised by a change of shape due to collapse of cell processes, and movement away from the other cell. (A-A”) EphB2 and ephrinB1 cells are both repelled following their interaction; (B-B”) EphB2-siSHIP2 cell has multiple cell processes and is not repelled, whereas ephrinB1 cell is repelled following their interaction; (C-C”) ephrinB1-siSHIP2 cell is not repelled, whereas EphB2 cell is repelled following their interaction. (D): quantitation of collapse responses from manual tracking of cells in time lapse movies (Movies 4, 5, 7). (E): Snapshots of time lapse movie (Movie 6) showing example of intercalation of EphB2-siSHIP2 cell (arrowhead) between ephrinB1 cells (asterisks). Scale bars: 20 μm.

Article Snippet: Antibodies used for Western blot analysis were to detect: EphB2 and GAPDH from Thermo Fisher (371700, MA5-15738, respectively); phosphorylated EphB2 from Abcam (ab61791); ephrinB1/2/3 from Santa Cruz (sc-910); FLAG tag from Sigma Aldrich (F3165); SHIP2 and phosphorylated SHIP2-Y1135 from Cell Signaling Technologies (#2839 and 5445, respectively).

Techniques: Quantitation Assay

Journal: bioRxiv

Article Title: Role of SHIP2 in cell repulsion regulated by Eph receptor and ephrin signaling

doi: 10.1101/872846

Figure Lengend Snippet: (A-C): immunostaining to detect pEphB2 (Y604) (white signal) and rab11, which marks intracellular vesicles (magenta signal), after 15 min of interaction. (A-A’) control EphB/ephrinB1 assay; (B-B’) after SHIP2 knockdown in EphB2 cells; (C-C’) after SHIP2 knockdown in ephrinB1 cells. EphB2 cells are labelled in green (examples indicated with white arrowheads) and ephrinB1 cells are unlabelled (magenta arrowheads). pEphB2 and rab11 are detected in retraction fibres and intracellular vesicles of EphB2 cells. Scale bars: 20 μm. (D) Western blot analysis of EphB2 phosphorylation (Y604) after forced interaction of EphB2 and ephrinB1 cells, with or without SHIP2 knockdown. (E) Quantitation of pEphB2 relative to EphB2 / ephrinB1 control at 5 min. There is no change in EphB2 phosphorylation following knockdown of SHIP2 in EphB2 cells, but it increases at 5 and 15 min following knockdown in ephrinB1 cells. This increase was found in two independent experiments, and the average is shown in (E).

Article Snippet: Antibodies used for Western blot analysis were to detect: EphB2 and GAPDH from Thermo Fisher (371700, MA5-15738, respectively); phosphorylated EphB2 from Abcam (ab61791); ephrinB1/2/3 from Santa Cruz (sc-910); FLAG tag from Sigma Aldrich (F3165); SHIP2 and phosphorylated SHIP2-Y1135 from Cell Signaling Technologies (#2839 and 5445, respectively).

Techniques: Immunostaining, Western Blot, Quantitation Assay

Journal: Neuron

Article Title: A sensitized IGF1 treatment restores corticospinal axon-dependent functions

doi: 10.1016/j.neuron.2017.07.037

Figure Lengend Snippet:

Article Snippet: KEY RESOURCES TABLE table ft1 table-wrap mode="anchored" t5 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Chicken monoclonal anti-GFP Abcam Cat#ab13970 Rabbit polyclonal anti-RFP Abcam Cat#ab34771 Rabbit anti-GFAP DAKO Cat# Z0334 Rabbit polyclonal anti-PKC gamma Santa Cruz Cat#sc211 Rabbit anti-5-HT Immunostar Cat#20080 Rabbit anti-IGFR Santa Cruz Cat#sc-712 Rabbit anti-pIGFRβ Cell Signaling Technology Cat#3024 Rabbit anti-pS6 Cell Signaling Technology Cat#4857 Rat anti-CD68 Bio-Rad Cat#MCA1957 Guinea pig-anti-Vglut1 Synaptic Systems Cat#135304 Goat-anti-ChAT Millipore Cat#AB144P Biological Samples N/A N/A N/A Chemicals, Peptides, and Recombinant Proteins Quipazine Sigma Cat#Q1004 SKF- 82197 Tocris Cat#1447 8-OH-DPAT Tocris Cat#0529 4-AP Sigma Cat#275875 4-AP-MeOH Santa Cruz Cat# sc-267247 Diptheria toxin Sigma Cat# D0564 Human recombinant IGF1 Peprotech Cat# 100–11 Human recombinant Osteopontin (OPN) Peprotech Cat# 120–35 Critical Commercial Assays N/A N/A N/A Deposited Data N/A N/A N/A Experimental Models: Cell Lines N/A N/A N/A Experimental Models: Organisms/Strains Mouse/C57Bl/6 Charles River Strain code#027 Mouse/Emx1Cre The Jackson Laboratory Jax#5628 Recombinant DNA AAV-PLAP Liu et al., 2010 Cat#N/A AAV-tdTomato Addgene Cat#59462 AAV-Cre This paper Cat#N/A AAV-OPN Duan et al., 2015 Cat#N/A AAV-IGF1 This paper Cat#N/A AAV-CaMKIIa-ChR2-mCherry Addgene Cat# 26975 AAV- Syn-ChR2-EYFP Addgene Cat#26973 Lenti-HiRet-GFP This paper Cat#N/A Lenti-HiRet-FLEX-DTR Jin, et al., 2015 Cat#N/A Sequence-Based Reagents N/A N/A N/A Software and Algorithms ImageJ2 NIH https://imagej.nih.gov/ij/index.html Prism 7.0 GraphPad Software, Inc. Cat# http://www.graphpad.com/ STATA 12 College station, TX, USA www.stata.com Other N/A N/A N/A Open in a separate window CONTACT FOR REAGENT AND RESOURCE SHARING Further information and requests for resources and reagents should be directed to and will be fulfilled by the Lead Contact, Dr. Zhigang He ( ude.dravrah.snerdlihc@eH.gnagihZ ).

Techniques: Recombinant, Sequencing, Software

Journal: Immunity

Article Title: Binding of IFIT1 by IFIT3 modulates protein stability and RNA binding specificity

doi: 10.1016/j.immuni.2018.01.014

Figure Lengend Snippet: Co-precipitation assays between IFIT1 and (A) IFIT2, (B) IFIT5, and (C–D) IFIT3. MBP-tagged IFIT proteins were incubated with amylose resin (lane 1) and unbound MBP-tagged protein was washed (lanes 2–5). Prebound MBP-tagged IFIT (lane 6) was incubated with untagged IFIT proteins (lane 7) prior to washes (lane 8–11) and final beads (lane 12). Bound protein was eluted with maltose-containing buffer (lane 13). Final beads and eluted samples (boxed) were assessed for binding. Data shown are representative of at least two independent experiments. (E) Structure and domain organization of IFIT1 bound to 5′ cap 0 RNA (left), including location of TPR regions. Cartoon representation of IFIT1 colored by subdomain (right). (F) Cartoon representation of IFIT1-FIT3-ap 0 RNA heterotrimeric complex structure: asymmetric unit shows IFIT1 molecule A (green), IFIT1 molecule B (cyan) bound to IFIT3CTD molecule C (magenta), IFIT3CTD molecule D (yellow), and 5′ cap 0 RNA (orange). See also Figure S1 and S2.

Article Snippet: Immunoblots were performed using a rabbit polyclonal anti-IFIT1 (Thermo Fisher), rabbit anti-GAPDH (Cell Signaling Technology), rabbit polyclonal anti-IFIT3 ( Fensterl et al., 2008 ), or mouse monoclonal anti-FLAG M2 (Sigma-Aldrich), followed by IRDye 680RD goat anti-rabbit or IRDye 800CW goat anti-mouse (LiCor).

Techniques: Incubation, Binding Assay

Journal: Immunity

Article Title: Binding of IFIT1 by IFIT3 modulates protein stability and RNA binding specificity

doi: 10.1016/j.immuni.2018.01.014

Figure Lengend Snippet: (A) The molecular interface between IFIT1 and IFIT3 is defined by HDX-MS. Differences in deuterium uptake induced by IFIT3 binding are displayed as a color gradient (see legend) and highlighted in the cartoon representation of IFIT1. (B) Comparison of deuterium uptake kinetic curves of IFIT1 (black) and IFIT1-IFIT3 (green). Representative ITC raw data and binding isotherm for (C) IFIT1 and IFIT3. Measured values are KD = 1.5 ± 2.0 nM, ΔH = − 1.3 ± 0.02 × 104 cal/mol, ΔS = − 2.19 cal/mol/deg, and n (no. of sites) = 0.93 ± 0.01. (D) IFIT1 and IFIT3ΔCTD. KD = not determined. (E) IFIT1 and IFIT3CTD. Measured values are KD = 1.4 ± 3.1 nM, ΔH = − 1.7 ± 0.06 × 104 cal/mol, ΔS = − 2.08 cal/mol/deg., and n (number of sites) = 0.97 ± 0.03. (F) IFIT1 and IFIT3CTD K426A-E439A-L445A-S451A-I453A-F457A mutant (IFIT3 CTD mut). KD = not determined. All experiments were performed at least four independent times. See also Figure S3 and S4.

Article Snippet: Immunoblots were performed using a rabbit polyclonal anti-IFIT1 (Thermo Fisher), rabbit anti-GAPDH (Cell Signaling Technology), rabbit polyclonal anti-IFIT3 ( Fensterl et al., 2008 ), or mouse monoclonal anti-FLAG M2 (Sigma-Aldrich), followed by IRDye 680RD goat anti-rabbit or IRDye 800CW goat anti-mouse (LiCor).

Techniques: Binding Assay, Mutagenesis

Journal: Immunity

Article Title: Binding of IFIT1 by IFIT3 modulates protein stability and RNA binding specificity

doi: 10.1016/j.immuni.2018.01.014

Figure Lengend Snippet: (A) Comparison of 5′-ppp RNA-bound IFIT5 (green) (PDB 4HOR) and IFIT1 bound to cap 0 RNA and IFIT3CTD (cyan; (primary PDB validation is provided), current structure). (B) Structural changes on IFIT1 upon IFIT3 binding as defined by HDX-MS. Differences in deuterium uptake induced by IFIT3 binding to IFIT1-cap 0 RNA complex are displayed as a color gradient (see legend) and highlighted in the cartoon representation of IFIT1. Data shown are representative of two independent experiments. Multiple peptides were identified by MS/MS, and only the ten highest abundant peptides were used in the analysis. (C) Quantitative filter binding assay for IFIT1 (solid) and IFIT1-IFIT3CTD (dotted) and cap 0 RNA (black), cap1 RNA (red), and 5′-ppp RNA (blue). The results are the average of at least three independent experiments. Error bars represent standard error of the mean (SEM). See also Figure S5.

Article Snippet: Immunoblots were performed using a rabbit polyclonal anti-IFIT1 (Thermo Fisher), rabbit anti-GAPDH (Cell Signaling Technology), rabbit polyclonal anti-IFIT3 ( Fensterl et al., 2008 ), or mouse monoclonal anti-FLAG M2 (Sigma-Aldrich), followed by IRDye 680RD goat anti-rabbit or IRDye 800CW goat anti-mouse (LiCor).

Techniques: Binding Assay, Tandem Mass Spectroscopy, Filter-binding Assay

Journal: Immunity

Article Title: Binding of IFIT1 by IFIT3 modulates protein stability and RNA binding specificity

doi: 10.1016/j.immuni.2018.01.014

Figure Lengend Snippet: IFIT3 gene edited (IFIT3mut/mut) 293T cells were trans-complemented with IFIT3 (IFIT3mut/mut + IFIT3), IFIT3 lacking the C-terminal tail (IFIT3mut/mut + IFIT3CTD), or firefly luciferase (IFIT3mut/mut + fluc). (A) 293T-IFIT3mut/mut + IFIT3 and 293T-IFIT3mut/mut + fluc cells were infected at an MOI of 5 with WNV WT or isogenic WNV NS5-E218A lacking 2′-O methylation of the cap structure of genomic RNA. Infection was measured 24 hpi by flow cytometry by staining for intracellular WNV E protein. The fraction of infected 293T-IFIT3mut/mut + IFIT3 cells relative to infected 293T-IFIT3mut/mut + fluc are shown for both viruses. Data are the mean of three independent experiments, and error bars represent SEM. (B–C) 293T-IFIT3mut/mut + IFIT3 and 293T-IFIT3mut/mut + fluc cells were infected with WNV WT or WNV NS5-E218A at an MOI of 0.001. Supernatant was collected at the indicated times after infection and analyzed by focus-forming assay. Data are the mean titers from six independent experiments, and errors bars represent SEM. (D) IFIT3mut/mut + IFIT3, IFIT3mut/mut + hIFIT3ΔCTD, IFIT3mut/mut + mIfit3CTD, and IFIT3mut/mut + fluc cells were stimulated with IFN-β and assessed for IFIT1 expression by Immunoblotting. One representative experiment of three is shown. (E–F) 293T cells expressing human IFIT1-flag under an inducible promoter (293T-IFIT1-doxy) were trans-complemented with IFIT3 or fluc and analyzed for IFIT1 expression (anti-flag tag) by Immunoblotting. (E) One of two representative Immunoblots is shown. (F) IFIT1 amounts were normalized to GAPDH in IFIT3 or fluc-transduced cells treated with doxycycline. Error bars represent SEM from two independent experiments. (G–H) 293T-IFIT1-doxy cells trans-complemented with IFIT3 or fluc were stimulated with doxycycline for 16 h at indicated concentrations, subsequently treated with 50 μM puromycin to arrest translation, and analyzed for IFIT1 (anti-flag tag) at the indicated time post-puromycin treatment. (G) One of three representative Immunoblots is shown. (H) IFIT1 protein amounts were quantified and normalized to the 0 h post-puromycin treatment. Error bars represent SEM from three independent experiments. The statistical significance shown is for the comparison of IFIT3 + 100 ng/ml doxy vs fluc + 100 ng/ml doxy; comparisons between other doses yielded similar results. (I-K) CRISPR-Cas9 gene editing was used to abolish IFIT1 expression from 293T-IFIT3mut/mut + IFIT3 and 293T-IFIT3mut/mut + fluc cells using two different IFIT1-targeting sgRNAs or as a control, scrambled sgRNAs. (I) IFIT1 expression following IFN-β stimulation was determined by Immunoblot. (J–K) 293T-IFIT3mut/mut + IFIT3 and 293T-IFIT3mut/mut + fluc cells that received IFIT1 or scrambled sgRNAs were infected with WNV WT or WNV NS5-E218A at an MOI of 5 (J) or WT or NS5-E218A ZIKV (Cambodia, 2010; strain FSS13025) at an MOI or 1 (K) and scored for infectivity at 24 (J) or 30 (K) hpi by flow cytometry. Infectivity was normalized to the fraction of infected 293T-IFIT3mut/mut + fluc cells that received scrambled sgRNAs. The mean relative infectivity from three (K) and five (J) independent experiments is shown, and error bars represent the SEM. In this Figure, statistical significance was determined using an ANOVA followed by Tukey’s (A, B, and C) or Sidak’s (H, and J–K) multiple comparisons tests (ns, not significant; *, P > 0.05; ***, P <0.001; ****, P < 0.0001). See also Figure S6.

Article Snippet: Immunoblots were performed using a rabbit polyclonal anti-IFIT1 (Thermo Fisher), rabbit anti-GAPDH (Cell Signaling Technology), rabbit polyclonal anti-IFIT3 ( Fensterl et al., 2008 ), or mouse monoclonal anti-FLAG M2 (Sigma-Aldrich), followed by IRDye 680RD goat anti-rabbit or IRDye 800CW goat anti-mouse (LiCor).

Techniques: Luciferase, Infection, Methylation, Flow Cytometry, Staining, Focus Forming Assay, Expressing, Western Blot, FLAG-tag, CRISPR

Journal: Immunity

Article Title: Binding of IFIT1 by IFIT3 modulates protein stability and RNA binding specificity

doi: 10.1016/j.immuni.2018.01.014

Figure Lengend Snippet: (A, B and E) 293T-IFIT1-doxy cells were transfected with plasmids encoding fluc, IFIT3, or the indicated IFIT3 mutants, and treated with doxycycline prior to infection with (A) WNV WT or (B and E) WNV NS5 E218A. Infection and transfection data were analyzed by flow cytometry 24 hpi (A and B, left panels; and also see Fig S6E) to determine the percentage of transfected cells that were positive for intracellular WNV E protein. Data are representative of three independent experiments, and error bars represent SEM. (A and B, right panels, and E) Data are normalized to infectivity of doxycycline-untreated cells for each independent transfection (IFIT3, IFIT3 mutants, or fluc) experiment. Errors bars represent the SEM, and data was pooled from three to six independent experiments for statistical analysis (below). (C–D) 293T-IFIT1-doxy cells were trans-complemented with IFIT3 or fluc and infected with (C) VEEV-TC83 (MOI of 1, 16 h) or (D) VSV (MOI of 1, 6 h). Left panels show the mean percentage of infected cells from one representative experiment, and error bars indicate the SEM from triplicate replicates. Right panels show data normalized to infectivity of doxycycline-untreated cells. Errors bars represent the SEM, and data was pooled from three independent experiments. In this Figure, statistical significance was determined using a t-test (A) or an ANOVA followed by Sidak’s (B, C, and D) or Dunnett’s (E) multiple comparisons tests (ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001). See also Figure S6.

Article Snippet: Immunoblots were performed using a rabbit polyclonal anti-IFIT1 (Thermo Fisher), rabbit anti-GAPDH (Cell Signaling Technology), rabbit polyclonal anti-IFIT3 ( Fensterl et al., 2008 ), or mouse monoclonal anti-FLAG M2 (Sigma-Aldrich), followed by IRDye 680RD goat anti-rabbit or IRDye 800CW goat anti-mouse (LiCor).

Techniques: Transfection, Infection, Flow Cytometry