Journal: The Journal of Cell Biology

Article Title: FMNL2 regulates dynamics of fascin in filopodia

doi: 10.1083/jcb.201906111

Figure Lengend Snippet: Disturbing the organization of F-actin alters filopodia and fascin movement. (A) Filopodia length in micrometers (left) and filopodia lifetime in seconds (right) of fascin knockdown HeLa cells expressing GFP-fascin. n = 22 cells, three independent experiments. (B) Filopodia length (normalized to DMSO control) and coverage (relative filopodia number/perimeter) was evaluated after long-time (left, n = 22, 28, or 25 cells) or short-time (right, n = 51, 28, or 22 cells) treatment with SMIFH2 and CK666. One-way ANOVA; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ns, not significant. (C) Percentage of fast-moving fascin was extracted from filopodia photoconversion data from HeLa cells treated with DMSO, SMIFH2, or CK666. Shown are data from three independent experiments ( n = 16, 18, or 11 filopodia respectively), one-way ANOVA. (D) Morphology of GFP-fascin–expressing HeLa cells embedded in soft 3D ECM after inhibition of formins (SMIFH2) and Arp2/3 complex (CK666). Images were captured using LLSM. Scale bar = 10 µm. (E) Inhibition of formins decreases pulling forces on ECM. Velocity magnitude of cell-induced bead displacement in 3D collagen matrix without ( n = 220 bead velocity vectors) and with inhibitor treatment. Analysis was performed as described in (SMIFH2, n = 140; CK666, n = 120; Jasplakinolide, n = 137; beads only, n = 60). **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. (F) Evaluation of fascin-S39A speed in comparison to WT fascin by FRAP. Filopodia were bleached in RFP-fascin WT (black) and RFP-fascin-S39A (green) expressing fascin knockdown HeLa cells, and FRAP intensities were determined over time. Representative of three independent experiments. n = 17 filopodia, two-way ANOVA with Bonferroni posttest; **, P ≤ 0.01. (G) Photoconversion of mEOS2-fascin in HeLa cells on 2D was performed to compare fascin recovery in lamellipodia and filopodia. n = 3 ROI measurements per cell area from 18 cells from three experiments. (H) Retention of fascin in lamellipodia is dependent on F-actin density. Small lamellipodial regions of DMSO control (black) and CK666-treated (magenta) GFP-fascin–expressing HeLa cells were photobleached, and FRAP was analyzed over time; n = 10 from one of three experiments. (I–K) Characterization of the CS-fascin. Filopodia length (I) and number (filopodia/perimeter; J) were measured for GFP-fascin–expressing cells and CS-fascin expressing cells. CS-fascin (tagged with both GFP and mScarlet) or GFP-fascin and mScarlet-fascin (K) were coexpressed in fascin knockdown HeLa cells; cells were fixed, and lifetime was measured by FLIM-FRET. Shown are data from two independent experiments ( n = 15, 16, or 12 cells). One-way ANOVA with Bonferroni posttest; ***, P ≤ 0.001; ****, P ≤ 0.0001. (L) CS-fascin–expressing cells were treated with the Arp2/3 inhibitor CK666 and imaged on a confocal microscope to measure ratiometric FRET. Bar graph shows FRET/donor (in %) for control ( n = 40 cells) and CK666 treatment ( n = 15 cells). One-way ANOVA with uncorrected Fisher posttest. (M) FRET signal of HeLa cells expressing CS-fascin from time binned datasets. Binning of time-lapse images shows increased FRET signal of CS-fascin both in filopodia and at the base of filopodia after combining 50 time-lapse frames. High FRET is shown in yellow/green, and low FRET is shown in dark blue/black. Imaging was performed by SIM; representative example is shown; scale bar = 5 µm. (N) Representative image of the GFP-fascin donor-only control for 3D FRET experiment shows the lack of filopodia and no FRET signal. Scale bar = 2 µm.

Article Snippet: The following compounds were used: CK666 and Jasplakinolide (Santa Cruz), SMIFH2 (Tocris), Cytochalasin D (Sigma-Aldrich), BIM (Sigma-Aldrich), phorbol 12,13-dibutyrate (Sigma-Aldrich), tetradecanoyl phorbol acetate (Sigma-Aldrich), and Kinesore (kindly provided by Mark Dodding, University of Bristol, UK).

Techniques: Knockdown, Expressing, Control, Inhibition, Comparison, Microscopy, Imaging

Journal: The Journal of Cell Biology

Article Title: FMNL2 regulates dynamics of fascin in filopodia

doi: 10.1083/jcb.201906111

Figure Lengend Snippet: Local F-actin organization regulates fascin movement into and out of filopodia. (A) Filopodia of mEOS2-fascin–expressing HeLa cells were photoconverted, and exit of photoconverted fascin was monitored over time using a confocal microscope at a 1-s frame rate. Upper panel, control cells (DMSO treated cells); middle panel, formin inhibition (SMIFH2-treated cells); bottom panel, Arp 2/3 complex inhibition (CK666-treated cells). Shown are pseudocolored ratios of photoconverted/unconverted intensities before (first column, −1 s) and after (shown are every 5 s) conversion. Scale bar = 5 µm. (B) Fascin movement out of filopodia. Photoconversion intensities plotted versus time for control and SMIFH2- and CK666-treated cells. Early and late phases were compared using two-way ANOVA with uncorrected Fisher’s posttest; shown are data ( n = 15, 24, or 17 cells, one ROI per cell, and two to four filopodia per ROI) from three independent experiments; *, P ≤ 0.05; **, P ≤ 0.01; ns, not significant. (C) Fascin t 1/2 values and mobility versus immobility (in %) extracted from monoexponential single-curve photoconversion fitting ( n = 15, 24, or 17 cells, one ROI per cell, and two to four filopodia per ROI). One-way ANOVA; *, P ≤ 0.05; **, P ≤ 0.01. (D) Fascin movement into filopodia. Filopodia of GFP-fascin–expressing HeLa were bleached (FRAP), and recovery intensities were plotted over time for control and SMIFH2- and CK666-treated cells. Early and late phases were compared using two-way ANOVA with uncorrected Fisher’s posttest; shown are data ( n = 18, 21, or 17 cells) from three independent experiments; *, P ≤ 0.05; **, P ≤ 0.01. (E) Fascin t 1/2 values and mobile versus immobile fractions extracted from monoexponential single-curve FRAP fitting ( n = 18, 21, or 17 cells). One-way ANOVA; *, P ≤ 0.05; ****, P ≤ 0.0001. (F) Small regions within single filopodia were bleached in control and SMIFH2- and CK666-treated cells and displayed as pseudocolored kymographs (space vs. time). Arrowheads point to the base or the tip of the filopodium. Scale bar, 1 μm. (G) Analysis of kymographs shown in F. Relative base to tip (left) and tip to base (right) speeds are shown. Data are from three independent experiments ( n = 18, 21, or 17 single filopodia from ≥15 individual cells per condition), one-way ANOVA; *, P ≤ 0.05; ***, P ≤ 0.001. (H) Interaction of GFP-fascin and RFP-actin measured in live cells by multiconfocal FLIM-FRET imaging. Examples of control cells (top panel) and SMIFH2 treated cells (bottom panel) are shown. Intensity images on the left and pseudocolored FLIM image on the right. Warm colors indicate FRET (low lifetime) and cool colors indicate no FRET (high lifetime). Scale bar = 2 µm. (I) Mean FRET efficiency (left, n = 16) and lifetime Lowess residuals (right, n = 28) of fascin–actin interaction of single filopodia measured by live FLIM in cells with and without SMIFH2 treatment. Student’s t test; *, P ≤ 0.05.

Article Snippet: The following compounds were used: CK666 and Jasplakinolide (Santa Cruz), SMIFH2 (Tocris), Cytochalasin D (Sigma-Aldrich), BIM (Sigma-Aldrich), phorbol 12,13-dibutyrate (Sigma-Aldrich), tetradecanoyl phorbol acetate (Sigma-Aldrich), and Kinesore (kindly provided by Mark Dodding, University of Bristol, UK).

Techniques: Expressing, Microscopy, Control, Inhibition, Imaging

Journal: Nature Communications

Article Title: A tripartite organelle platform links growth factor receptor signaling to mitochondrial metabolism

doi: 10.1038/s41467-024-49543-z

Figure Lengend Snippet: a CD147 internalization was monitored in HeLa cells subjected to N-WASP-KD, RTN3-KD or treatment with CK666 and stimulated with high dose EGF. Left, representative IF images, CD147 (green), EGF (red), DAPI (blue), Bar, 10 µm. Right upper, quantification of relative CD147 fluorescence intensity expressed as % of control. N = number of cells: Control/N = 338 ( n = 4); RTN3 = KD/N = 158, NWASP-KD/N = 237, CK666/N = 250 ( n = 3). Right lower, Alexa647-Tf internalization was monitored for 8 min at 37 °C in HeLa cells treated as above. The data reported for RTN3 KD are the same as displayed in Fig. . N=number of cells: Control/N = 246 ( n = 6); RTN3-KD/N = 185, NWASP-KD/N = 276, CK666/N = 295 ( n = 3). Box plots are defined as in Fig. . b , c EM morphometric analysis of the length of EGFR gold-positive TIs ( b ) and CCPs ( c ) in HeLa cells subjected to dynamin KD (Dyn-KD) and/or treated with CK666. Data are expressed as Fig. , mean ± SD; N = cell profiles. Control/N = 10, CK666/N = 10, Dyn-KD/N = 9, Dyn-KD/CK666/N = 10 ( b ); Control/N = 10, CK666/N = 10, Dyn-KD/N = 9, Dyn-KD/CK666/N = 10 (c); a representative experiment of n = 2 is shown. d Ratiometric analysis of membrane proximal actin (MPA) density. HeLa cells were subjected or not to RTN3 KD, followed by MPAct-mCherry and YFP-CaaX co-transfection, and stimulated with high or low EGF or left unstimulated (-EGF). Left, two representative images of the ratiometric analysis of MPA density (MPAct signal normalized over the CaaX PM marker). Bar, 10 μm. e Right, mean raw integrated fluorescence intensity ±SD is reported as a percentage relative to control cells. N = number of cells: -EGF/N = 424, LowEGF/N = 495, HighEGF/N = 568 ( n = 7). f , g Ratiometric analysis of MPA density as in d , in HeLa cells subjected to the indicated KDs ( f ) or inhibitor treatments ( g ) and stimulated with high dose EGF. Data are expressed as mean ± SD. Inhibitors were present during the stimulation. N = number of cells: ( f ) -EGF/N = 316/n = 5, Control/N = 356/n = 5, RTN3-KD/N = 409/n = 5, RTN4-KD/N = 311/n = 4, NWASP-KD/N = 370/n = 4; ( g ) Control/N = 306, CK666/N = 228, XeC/N = 231, MCUi11/N = 232, OMY/N = 189 ( n = 3). Exact p -values (Each pair Student’s t test, two-tailed) are shown in all panels; ns, not significant; n=biological replicates. Source data are provided as a Source Data file.

Article Snippet: Oligomycin (OMY), histamine, and succinate were from Sigma, xestospongin C (XeC) was from abcam, CK666 and bongkrekic acid were from Merck Life Technology S.R.L., MCUi11 was from CliniSciences and Gefitinib was from Merck Life Technology S.R.L., Ciliobrevin D was from TargetMol (T14965).

Techniques: Fluorescence, Control, Membrane, Cotransfection, Marker, Two Tailed Test

Journal: Nature Communications

Article Title: A tripartite organelle platform links growth factor receptor signaling to mitochondrial metabolism

doi: 10.1038/s41467-024-49543-z

Figure Lengend Snippet: a CD147 internalization in HaCaT cells subjected to the indicated KDs or mock transfection (Ctrl). Internalized CD147 was measured by IF after stimulation with high dose Alexa647-EGF for 12 min at 37 °C, as described in Fig. . Mean integrated fluorescence intensity is reported as a percentage of the control cells. N = number of cells: Control/N = 301/n = 6, RTN3-KD/N = 97/n = 3, RTN4-KD/N = 179/n = 5, MCU-KD/N = 221/n = 4, IP3R-KD/N = 210/n = 4. b HaCaT cells expressing the Ca 2+ sensor PM-GCaMP6f were stimulated with high or low EGF. Left, Representative single-cell Ca 2+ response curves presented as the ratio of fluorescence emission at 490/406 nm. Right, AUC of the Ca 2+ response. N=number of cells: High EGF/N = 17, Low EGF/N = 20 ( n = 2). c Ca 2+ response in HaCaT cells subjected to RTN3 or RTN4 KD or to mock transfection (Ctrl) and stimulated with high EGF dose as in b . AUC is shown. N = number of cells: Control/N = 18, RTN3-KD/N = 25, RTN4-KD/N = 14. A representative experiment of three independent replicates is shown. d ΔΨm in HaCaT cells stimulated or not with low or high EGF, measured using TMRM. Total peak area is reported. N=number of cells: -EGF/N = 206, Low EGF/N = 253, High EGF/N = 327. A representative experiment of n = 2 is shown. e Wound healing in sub-confluent HaCaT cells stimulated or not with low or high EGF was monitored by time-lapse video microscopy. Left, representative images at the indicated time points. Middle, distance covered by cells in the different conditions corrected to the baseline at t = 0. Right, wound closure velocity (μm/h) at t = 24 h is shown. N=number of cells: -EGF/N = 15, Low EGF/N = 23, HighEGF/N = 34 ( n = 3). f Wound closure velocity (μm/h) in sub-confluent HaCaT cells stimulated with high EGF dose and subjected to RTN3 KD or the indicated treatments for 24 h. N = number of cells: Control/N = 98/n = 6, DMSO/N = 24/n = ), RTN3-KD/N = 40/n = 2, OMY/N = 44/n = 3, MCUi11/N = 22/n = 2, CK666/N = 28/n = 2, XeC/N = 30/n = 2). g Schematic showing the crosstalk between EGFR-NCE, ER, and mitochondria in controlling EGFR endocytosis, receptor fate and cell migration. Box plots in all panels are defined as in Fig. . Exact p -values (each pair Student’s t -test, two-tailed) are shown; ns not significant, n biological replicates. Source data are provided as a Source Data file.

Article Snippet: Oligomycin (OMY), histamine, and succinate were from Sigma, xestospongin C (XeC) was from abcam, CK666 and bongkrekic acid were from Merck Life Technology S.R.L., MCUi11 was from CliniSciences and Gefitinib was from Merck Life Technology S.R.L., Ciliobrevin D was from TargetMol (T14965).

Techniques: Transfection, Fluorescence, Control, Expressing, Single Cell, Microscopy, Migration, Two Tailed Test

Journal: The Journal of Cell Biology

Article Title: Type V myosin focuses the polarisome and shapes the tip of yeast cells

doi: 10.1083/jcb.202006193

Figure Lengend Snippet: Actin is required for polarisome focusing. (A) myo2-66 cells carrying an empty plasmid or expressing MYO2 or myo2 RD from the genomic URA3 locus were spotted in 10-fold serial dilutions on SD-Ura − plates containing 70 µM methionine and incubated for 3 d at either 25°C (upper panel) or 30°C (lower panel). (B) myo2-66 cells carrying an empty plasmid or expressing MYO2 or myo2 RD from the genomic URA3 locus and coexpressing Spa2-mCherry were analyzed by fluorescence microscopy at either 27°C (left panel) or 31°C (right panel). Insets show the DIC image of the same cell in reduced magnification. (C) Intensity profiles of the Spa2-mCherry signals of the bud circumference of the cells in B at 27°C. (D) The polarisome distribution in the bud of the cells from B was classified as focused (gray), cortically spread (spread, blue), or diffuse (black; 27°C: n empty = 226, n MYO2 = 232, n myo2RD = 225; 31°C: n MYO2 = 223, n myo2RD = 224). (E) Left: Fluorescence microscopy of WT, bni1Δ , or bni1 Δ 1240–end cells expressing Spa2-GFP. Right: Intensity profiles of Spa2-GFP of the bud circumference of the cells from the left panel. (F) The polarisome distribution of the cells from E were classified as either focused (gray) or cortically spread (spread, blue; n WT = 234, n bni1Δ = 234, n bni1Δ1240–end = 227). (G) The actin cytoskeleton of WT (upper panel) or myo2 RD cells (lower panel) was stained with Alexa Fluor 488–phalloidin in the absence (left) or presence (right) of CK-666. All scale bars: 5 µm.

Article Snippet: Actin patches were removed by addition of 100 µM CK-666 (Merck) to the cell medium at 30°C, 10 min before cells were fixed.

Techniques: Plasmid Preparation, Expressing, Incubation, Fluorescence, Microscopy, Staining

Journal: Microorganisms

Article Title: Herpes Simplex Virus Type 1 Infection Induces the Formation of Tunneling Nanotubes

doi: 10.3390/microorganisms11081916

Figure Lengend Snippet: Fluorescence microscopy images showed the effect of CK666 on the formation of TNTs. ( A , B ) HCECs and Vero cells were stained with phalloidin green to visualize the nanotubes. Three images (per experiment) were taken per condition, with −100 cells examined, and the number of TNTs was counted. ( C ) Graph shows a significant reduction in the number of TNTs in the presence of CK666 (50 μM). Data are presented as mean ± SD. Student’s t -test was performed to compare between the groups, and the following significance values were considered: * p < 0.05, *** p < 0.001. ( D ) Cell viability of human corneal epithelial cells and Vero cells in the presence of CK666. Cells were treated with different concentrations of inhibitors, 0.1–100 μM, for 24 h, and the cell viability was measured by MTT assays. Only 100 μM was found to be toxic in two types of cells. Data shown are of three replicates.

Article Snippet: The 10 4 HCECs and Vero cells in 96-wells plate were treated with various concentrations of CK666 (AdooQ Bioscience LLC., Irvine, CA, USA) (0, 0.1, 1, 5, 10, 20, 50, 100 μM) and 0.1% dimethyl sulfoxide (DMSO, as a drugs vehicle) (Sigma, Cream Ridge, NJ, USA) for 24 h. After incubation with CK666 and DMSO, cells were incubated with 5 mg/mL concentration of MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (Beyotime Biotechnology Co., Ltd., Haimen, China) reagent in media for 4 h. Then, cells were incubated with Formazan solvent for 4 h and absorbance recorded using Multiskan Spectrum at 570 nm (Multiskan GO, Thermo Scientific, Waltham, MA, USA).

Techniques: Fluorescence, Microscopy, Staining

Journal: Microorganisms

Article Title: Herpes Simplex Virus Type 1 Infection Induces the Formation of Tunneling Nanotubes

doi: 10.3390/microorganisms11081916

Figure Lengend Snippet: CK666 affects the transmission of HSV-1. Confluent monolayers of cell plates in 24-well culture were infected with HSV-1 for 1 h, then treated with/without 50 μM CK666 for 24 h in HCECs ( A ) and Vero cells ( B ) and processed for the plaque assay. ( C ) Graph shows a reduction in the number of plaques in HCECs and Vero cells treated with CK666 compared to control cells. ( D ) HCECs and Veros were infected with wild-type HSV-1 (MOI of 1), and the effect of the addition of 50 μM CK666 on the HSV-1 DNA level in HCECs and Vero cells was quantified with qPCR. Data represent mean ± SD of triplicate assays. Student’s t -test was used to compare between the groups. ** p < 0.01, *** p < 0.001.

Article Snippet: The 10 4 HCECs and Vero cells in 96-wells plate were treated with various concentrations of CK666 (AdooQ Bioscience LLC., Irvine, CA, USA) (0, 0.1, 1, 5, 10, 20, 50, 100 μM) and 0.1% dimethyl sulfoxide (DMSO, as a drugs vehicle) (Sigma, Cream Ridge, NJ, USA) for 24 h. After incubation with CK666 and DMSO, cells were incubated with 5 mg/mL concentration of MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (Beyotime Biotechnology Co., Ltd., Haimen, China) reagent in media for 4 h. Then, cells were incubated with Formazan solvent for 4 h and absorbance recorded using Multiskan Spectrum at 570 nm (Multiskan GO, Thermo Scientific, Waltham, MA, USA).

Techniques: Transmission Assay, Infection, Plaque Assay, Control

Journal: bioRxiv

Article Title: Keratins and Plakin family cytolinker proteins control the length of epithelial microridge protrusions

doi: 10.1101/2020.02.18.954933

Figure Lengend Snippet: (A) SIM optical sections of a Krt17-GFP[BAC]-expressing zebrafish periderm cell at 48 hpf. Cartoon shows the relative location of Apical (A) and Medial (M) optical sections. (B) Oblique optical section through a cell with endogenously-tagged Krt17. This section shows both the microridge-like pattern at the apical surface of the cell (arrow), and the filamentous pattern deeper in the cell (arrowhead). (C) Projections and orthogonal views of SIM images of Krt17-GFP[BAC]- and Lifeact-Ruby-expressing cells at the indicated developmental stages. White boxes, regions of magnification in zoom panels. Orthogonal views (XZ) at 19hpf show Krt17 is not in pegs, but at 48 hpf localizes alongside Actin in microridges. (D) Dot and box plot of average microridge length per cell in cells over-expressing Krt17-GFP[BAC] at 48hpf. *p<0.05, Wilcoxon rank-sum test. n=24 cells from 4 fish. Blue line shows the median average microridge length per cell in WT animals (see ). (E) Dot and box plot of average microridge length per cell in cells at 48hpf after 30min treatment with DMSO or CK666. ***p<0.005, Wilcoxon rank-sum test. n=11-12 cells from 3 fish. (F) Dot and box plot of microridge and Keratin overlap coefficients, comparing the colocalization of each protein at 0 min with its localization after 30 min DMSO or CK666 treatment. **p<0.01, ***p<0.005, Wilcoxon rank-sum test. n=11-12 cells from 3. (G) Krt17-GFP[BAC]- and Lifeact-Ruby-expressing cells at 48hpf treated with CK666 for 30 minutes. Arrows show that the F-actin microridge pattern was disrupted after 30min of treatment. Arrowheads show that the Krt17 microridge pattern was retained. See for images of control DMSO treatments. (H) GFP-PH-PLC and Krt17-mRuby[BAC] in periderm cells treated with CK666 for 30 minutes at 48hpf. GFP-PH-PLC was expressed by crossing BAC(ΔNp63:Gal4FF) la213 to UAS:GFP-PH-PLC, and Krt17-mRuby[BAC] was expressed by transient transgenesis. Arrowheads show that membrane protrusions and the Krt17 microridge pattern were maintained. See for images of control DMSO treatments, and orthogonal views. Scale bars:10µm (A-C, G-H, blue line) and 1µm (C, white line).

Article Snippet: CK666 (Fisher Scientific) was dissolved in DMSO.

Techniques: Expressing

Journal: bioRxiv

Article Title: Keratins and Plakin family cytolinker proteins control the length of epithelial microridge protrusions

doi: 10.1101/2020.02.18.954933

Figure Lengend Snippet: (A) Krt17-GFP[BAC]- and Lifeact-Ruby-expressing cells in 48hpf fish at time 0 and 30 minutes after DMSO or CK666 treatment. Arrows show that the F-actin microridge pattern was disrupted after 30 min of CK666 treatment. Arrowheads show that the Keratin microridge pattern was retained after 30min of CK666 or DMSO treatment. Image of CK666 treatment is replicated from , for comparison. (B) GFP-PH-PLC (membrane) and Lifeact-Ruby in 48 hpf periderm cells at time 0 and 30min after treatment with DMSO or CK666. BAC(ΔNp63:Gal4FF) la213 to UAS:GFP-PH-PLC, and Krt5:Lifeact-Ruby was expressed by transient transgenesis. Arrows show that the membrane and Actin microridge pattern was disrupted after 30min of CK666 treatment. Arrowheads show that the membrane and Actin microridge pattern was retained in DMSO controls. (C) Projection and orthogonal views of GFP-PH-PLC and Krt17-mRuby[BAC] in 48hpf periderm cells at time 0 and 30min after treatment with DMSO or CK666. BAC(ΔNp63:Gal4FF) la213 to UAS:GFP-PH-PLC, and Krt17-mRuby[BAC] was expressed by transient transgenesis. Arrows show that the membrane and Krt17 microridge patterns were retained after 30min DMSO or CK666 treatment. Orthogonal views (XZ) show that Krt17 preserved the protrusive membrane structure after 30min CK666 treatment. Image of CK666 treatment is replicated from , for comparison. Scale bars: 10µm (A-C) and 1µm (Orthogonal images in C).

Article Snippet: CK666 (Fisher Scientific) was dissolved in DMSO.

Techniques: Expressing

Journal: EMBO Reports

Article Title: Nesprin‐2 accumulates at the front of the nucleus during confined cell migration

doi: 10.15252/embr.201949910

Figure Lengend Snippet: Reagents and Tools table

Article Snippet: CK666 (VWR) was dissolved to a stock concentration of 250 mM in dry DMSO and used at a working concentration of 0.25 mM.

Techniques: Sequencing, Stable Transfection, Expressing, Construct, Mutagenesis, Recombinant, Plasmid Preparation, Imaging, Software