Journal: bioRxiv

Article Title: Two independent translocation modes drive neural stem cell dissemination into the human fetal cortex

doi: 10.1101/2025.01.08.631865

Figure Lengend Snippet: (A) Quantification of IST amplitude in bRG cells following treatment with DMSO, nocodazole (1 μ M) or blebbistatin (10 μ M) in cortical organoids (N=3 organoid batches, 329 bRG cells, weeks 9-12). (B) Live imaging of mitotic human bRG cells expressing control, DYNC1H1 or LIS1 shRNA constructs in human cortical organoids (week 8-11). shRNA plasmids co-express GFP. (C) Live imaging of mitotic human bRG cells expressing control, DYNC1H1 or LIS1 shRNA constructs in human fetal tissue (pcw 16-20). (D) Live imaging of in vitro interphasic human bRG cells expressing control or LIS1 shRNA constructs. (E) Quantification of IST amplitude in in vitro interphasic human bRG cells expressing control or LIS1 shRNA constructs (N=3 experiments, 520 bRG cells). (F) Quantification of IST amplitude in in vitro interphasic human bRG cells expressing control or KASH constructs, in the presence of DMSO or blebbistatin (10 μ M) (N=3 experiments, 1198 bRG cells). Yellow arrowheads indicate bRG cell soma, and green and red arrowheads indicate daughter cells. Data are presented as mean values +/− SD. Scale bar = 20 µm. All live imaging montages are in hours:minutes. **p<0,01; ***p<0,001, ns: non-significant by two-tailed unpaired t-tests.

Article Snippet: The following plasmids were used in this study: MSCV-IRES-GFP (Tannishtha Reya, Addgene 20672); VSVG (a gift from P. Benaroch), Human EZR shRNA (TF308420, Origene), Human STK10 shRNA (TF320540, Origene), Human SLK shRNA (TG320620, Origene), Human DYNC1H1 shRNA (TL313335, Origene), Human RDX shRNA (TL309884, Origene), Human MSN shRNA (TL311375, Origene), Human ECT2 shRNA (TL304854, Origene), Human VIM shRNA (TL308419, Origene), Human PAFAH1B1 (LIS1) shRNA (TL310628, Origene), Dominant Negative KASH.

Techniques: Imaging, Expressing, Control, shRNA, Construct, In Vitro, Two Tailed Test

Journal: bioRxiv

Article Title: Two independent translocation modes drive neural stem cell dissemination into the human fetal cortex

doi: 10.1101/2025.01.08.631865

Figure Lengend Snippet: (A) Live imaging of interphasic human bRG cells expressing control, DYNC1H1 or LIS1 shRNA constructs in human cortical organoids (week 8-11). shRNA plasmids co-express GFP. (B) Live imaging of interphasic human bRG cells expressing control, DYNC1H1 or LIS1 shRNA constructs in human fetal tissue (pcw 16-18). (C) Quantification of IST amplitude in human bRG cells expressing control, DYNC1H1 or LIS1 shRNA constructs in human cortical organoids (N=3 organoid batches, 899 bRG cells, week 8-11). Two independent shRNA plasmids were used for each knockdown. (D) Quantification of MST amplitude in human bRG cells expressing control, DYNC1H1 or LIS1 shRNA constructs in human cortical organoids (N=3 organoid batches, 899 bRG cells, week 8-11). (E) Quantification of IST amplitude in human bRG cells expressing control, DYNC1H1 or LIS1 shRNA constructs in human fetal tissue (N=3 fetal samples, 385 bRG cells, pcw 16-20). (F) Quantification of MST amplitude in human bRG cells expressing control, DYNC1H1 or LIS1 shRNA constructs in human fetal tissue (N=3 fetal samples, 385 bRG cells, pcw 16-20). (G) Live imaging of interphasic human bRG cells expressing GFP in control cortical organoids and two different patient-derived LIS1-mutated organoids (week 8-11). (H) Quantification of IST amplitude in control cortical organoids and two different patient-derived LIS1-mutated organoids (N=3 organoid batches, 397 bRG cells, week 8-11). (I) Quantification of MST amplitude in control cortical organoids and two different patient-derived LIS1-mutated organoids (N=3 organoid batches, 397 bRG cells, week 8-11). (J) Immunostaining for SOX2 and Nesprin-2 in cortical organoids expressing GFP or the KASH dominant negative together with GFP (week 9). Red arrows indicate nuclear envelope of construct-expressing cells. (K) Live imaging of interphasic human bRG cells expressing control or KASH constructs in human cortical organoids (week 8). KASH plasmid co-expresses GFP. (L) Live imaging of interphasic human bRG cells expressing control or KASH constructs in human fetal tissue (pcw 16). KASH plasmid co-expresses GFP. (M) Quantification of IST amplitude in human bRG cells expressing control or KASH constructs in human cortical organoids (N=3 organoid batches, weeks 8-11, 201 bRG cells). (N) Quantification of MST amplitude in human bRG cells expressing control or KASH constructs in human cortical organoids (N=3 organoid batches, weeks 8-11, 201 bRG cells). (O) Quantification of IST amplitude in human bRG cells expressing control or KASH constructs in human fetal tissue (N=2 fetal samples, pcw 16-18, 40 bRG cells). (P) Quantification of MST amplitude in human bRG cells expressing control or KASH constructs in human fetal tissue (N=2 fetal samples, pcw 16-18, 40 bRG cells). Yellow arrowheads indicate bRG cell soma, and green and red arrowheads indicate daughter cells. Data are presented as mean values +/− SD. Scale bar = 20 µm. All live imaging montages are in hours:minutes. **p<0,01; ****p<0,0001, ns: non-significant by two-tailed unpaired t-tests.

Article Snippet: The following plasmids were used in this study: MSCV-IRES-GFP (Tannishtha Reya, Addgene 20672); VSVG (a gift from P. Benaroch), Human EZR shRNA (TF308420, Origene), Human STK10 shRNA (TF320540, Origene), Human SLK shRNA (TG320620, Origene), Human DYNC1H1 shRNA (TL313335, Origene), Human RDX shRNA (TL309884, Origene), Human MSN shRNA (TL311375, Origene), Human ECT2 shRNA (TL304854, Origene), Human VIM shRNA (TL308419, Origene), Human PAFAH1B1 (LIS1) shRNA (TL310628, Origene), Dominant Negative KASH.

Techniques: Imaging, Expressing, Control, shRNA, Construct, Knockdown, Derivative Assay, Immunostaining, Dominant Negative Mutation, Plasmid Preparation, Two Tailed Test

Journal: bioRxiv

Article Title: Two independent translocation modes drive neural stem cell dissemination into the human fetal cortex

doi: 10.1101/2025.01.08.631865

Figure Lengend Snippet: (A) Immunostaining for SOX2 and p-ERM in mitotic bRG cell from week 8 cortical organoid. (B) Live imaging of in vitro mitotic human bRG cells expressing control, Moesin, Radixin, Ezrin or Vimentin shRNA constructs. (C) Quantification of IST amplitude in in vitro interphasic human bRG cells expressing control, Moesin, Radixin, Ezrin or Vimentin shRNA constructs. Two independent shRNA plasmids were used for each knockdown (N=7 control samples and N=3 sample per shRNA condition, 522 bRG cells). (D) Live imaging of mitotic human bRG cells expressing control, Moesin or Vimentin shRNA constructs in human cortical organoids (week 9). (E) Live imaging of mitotic human bRG cells expressing control, Moesin or Vimentin shRNA constructs in human fetal tissue (pcw 20). (F) Quantification of MST amplitude in human bRG cells expressing control, Moesin or Vimentin shRNA constructs in human cortical organoids (N=3 organoid batches, 453 bRG cells, weeks 8-12). (G) Quantification of IST amplitude in human bRG cells expressing control, Moesin or Vimentin shRNA constructs in human cortical organoids (N=3 organoid batches, 453 bRG cells, weeks 8-12). (H) Quantification of MST amplitude in human bRG cells expressing expressing control, Moesin or Vimentin shRNA constructs in human fetal tissue (N=3 fetal samples, 529 bRG cells, pcw 14-20). (I) Quantification of IST amplitude in human bRG cells expressing expressing control, Moesin or Vimentin shRNA constructs in human fetal tissue N=3 fetal samples, 529 bRG cells, pcw 14-20). (J) Schematic representation of the molecular mechanisms driving IST and MST. Yellow arrowheads indicate bRG cell soma, and green and red arrowheads indicate daughter cells. Data are presented as mean values +/− SD. Scale bar = 20 µm. All live imaging montages are in hours:minutes. *p<0,05; **p<0,01, ns: non-significant by two-tailed unpaired t-tests.

Article Snippet: The following plasmids were used in this study: MSCV-IRES-GFP (Tannishtha Reya, Addgene 20672); VSVG (a gift from P. Benaroch), Human EZR shRNA (TF308420, Origene), Human STK10 shRNA (TF320540, Origene), Human SLK shRNA (TG320620, Origene), Human DYNC1H1 shRNA (TL313335, Origene), Human RDX shRNA (TL309884, Origene), Human MSN shRNA (TL311375, Origene), Human ECT2 shRNA (TL304854, Origene), Human VIM shRNA (TL308419, Origene), Human PAFAH1B1 (LIS1) shRNA (TL310628, Origene), Dominant Negative KASH.

Techniques: Immunostaining, Imaging, In Vitro, Expressing, Control, shRNA, Construct, Knockdown, Two Tailed Test

Journal: bioRxiv

Article Title: Two independent translocation modes drive neural stem cell dissemination into the human fetal cortex

doi: 10.1101/2025.01.08.631865

Figure Lengend Snippet: (A) Live imaging of an in vitro GBM cell (line U3123) performing IST. (B) Live imaging of an in vitro GBM cell (line U3123) performing MST. (C) Quantification of the fraction of cells performing IST, in 9 GBM lines and compared to in vitro bRG cells (N=3 replicates per line, 1130 cells). (D) Quantification of the fraction of cells performing MST, in 9 GBM lines and compared to in vitro bRG cells (N=3 replicates per line, 1130 cells). (E) Immunostaining for bRG markers SOX2 and HOPX in U3123 GBM line. (F) Live imaging of U3123 GBM line during interphase, treated with DMSO, nocodazole (1 μ M) or blebbistatin (10 μ M). (G) Quantification of IST amplitude following treatment with DMSO, nocodazole (1 μ M) or blebbistatin (10 μ M) in U3123 GBM line (N=3 experiments, 274 GBM cells). (H) Live imaging of U3123 GBM line during mitosis, treated with DMSO, nocodazole (1 μ M) or blebbistatin (10 μ M). (I) Quantification of MST amplitude following treatment with DMSO, nocodazole (1 μ M) or blebbistatin (10 μ M) in U3123 GBM line (N=3 experiments, 274 GBM cells). (J) Live imaging of interphasic U3123 GBM cells expressing control, LIS1 shRNA or KASH dominant negative constructs. (K) Quantification of IST amplitude in U3123 GBM cells expressing control or LIS1 shRNA constructs. (N=3 experiments, 359 GBM cells). (L) Quantification of IST amplitude in U3123 GBM cells expressing control or KASH dominant negative constructs (N=3 experiments, 300 GBM cells). (M) Live imaging of mitotic U3123 GBM cells expressing control, Moesin or Vimentin shRNA constructs. (N) Quantification of MST amplitude U3123 GBM cells expressing control, Moesin or Vimentin shRNA (N=3 experiments, 149 GBM cells). Yellow arrowheads indicate bRG cell soma, and green and red arrowheads indicate daughter cells. Data are presented as mean values +/− SD. Scale bar = 20 µm. All live imaging montages are in hours:minutes. **p<0,01; ***p<0,001; ****p<0,0001, ns: non-significant by two-tailed unpaired t-tests.

Article Snippet: The following plasmids were used in this study: MSCV-IRES-GFP (Tannishtha Reya, Addgene 20672); VSVG (a gift from P. Benaroch), Human EZR shRNA (TF308420, Origene), Human STK10 shRNA (TF320540, Origene), Human SLK shRNA (TG320620, Origene), Human DYNC1H1 shRNA (TL313335, Origene), Human RDX shRNA (TL309884, Origene), Human MSN shRNA (TL311375, Origene), Human ECT2 shRNA (TL304854, Origene), Human VIM shRNA (TL308419, Origene), Human PAFAH1B1 (LIS1) shRNA (TL310628, Origene), Dominant Negative KASH.

Techniques: Imaging, In Vitro, Immunostaining, Expressing, Control, shRNA, Dominant Negative Mutation, Construct, Two Tailed Test

Journal: bioRxiv

Article Title: Myeloid CD209 impairs T-cell activation by inducing ICAM-2–ERM–dependent cortical stiffening

doi: 10.64898/2026.01.19.699861

Figure Lengend Snippet: a. Jurkat cells were treated with plate-coated Fc or hCD209-Fc for the indicated times, followed by immunoblotting of the indicated proteins. b. Control Jurkat cells or ICAM-2 KO Jurkat cells were treated with plate-coated hCD209-Fc for 0, 15 or 30 min, followed by immunoblot analysis of the indicated proteins. c. Jurkat cells were treated with plate-coated Fc or hCD209-Fc for 30 mins. After fixation, immunofluorescence staining was performed on p-ERM (red), phalloidin (green), and DAPI (blue) to observe the colocalization of p-ERM and phalloidin. d-e. Jurkat cells were treated with plate-coated Fc or hCD209-Fc for 30 min, followed by confocal microscopy analysis. A representative image of x-z through the equatorial plane of the cells is shown ( d ). The deformability index (x/z) is shown as a bar graph ( e , n=54/56). f-g. Control ( f , n=21/21) Jurkat cells or ICAM-2 KO ( g ,n=33/33) Jurkat cells were treated as described in d , and the deformability index (x/z) is shown. h-k. Wild-type Jurkat cells ( h ), control Jurkat cells, ICAM-2 KO Jurkat cells ( i ) or Jurkat cells overexpressing ezrin-WT, ezrin-T567A or ezrin-T567D ( k ) were treated with Fc or hCD209-Fc for 30 minutes and then incubated with Raji cells stimulated with SEE pulses at 37 °C for 15 minutes, The percentages of conjugates between Jurkat cells and Raji cells were analysed by flow cytometry ( h-i, k ). Lysates of Jurkat cells overexpressing ezrin-WT, ezrin-T567A or ezrin-T567D were immunoblotted for different proteins ( j ). l. Jurkat cells were treated with plate-coated CD209-Fc recombinant proteins for the indicated times. The cell lysates were immunoprecipitated with an anti-ezrin antibody, followed by immunoblot analysis of the indicated proteins. m. Jurkat cells were left untreated or pretreated with Y27632 (20 μg/mL) or ROCK-IN-1 (20 μg/mL) for 4 hours. The cells treated with the inhibitor were subsequently stimulated with hCD209-Fc protein for the indicated times, followed by immunoblot analysis of the indicated proteins. n. Jurkat cells were pretreated with Y27632 (20 μg/mL) or ROCK-IN-1 (20 μg/mL) for 4 hours. The cells treated with the inhibitor were subsequently stimulated with Fc or hCD209-Fc protein for 30 minutes and then incubated with Raji cells stimulated with SEE pulses at 37 °C for 15 minutes. The percentages of conjugates between Jurkat cells and Raji cells were analysed by flow cytometry. *: P<0.05; **: P<0.01; ***: P<0.005; ****: P<0.001 based on two-way ANOVA ( h-i, k and n ) and two-tailed unpaired t test ( e-g ). All error bars represent the S.E.M. of technical replicates ( e-i, k and n ), and the data are representative of two independent experiments ( a-c, h-n ). The data from three independent replications were pooled together in d-g .

Article Snippet: Antibodies against p-ERK (clone 12D4, (Santa Cruz Biotechnology Cat# sc-81492, RRID:AB_1125801)), HSP90 (clone AC-16, (Santa Cruz Biotechnology Cat# sc-101494, RRID:AB_1124018)), actin (clone C-2, (Santa Cruz Biotechnology Cat# sc-8432, RRID:AB_626630)), GAPDH (clone A-3, (Santa Cruz Biotechnology Cat# sc-137179, RRID:AB_2232048)), ERK (G-8, (Santa Cruz Biotechnology Cat# sc-271269, RRID:AB_10611091)), and ezrin (6A84, (Santa Cruz Biotechnology Cat# sc-71082, RRID:AB_1122712)) were purchased from SANTA CRUZ BIOTECHNOLOGY.

Techniques: Western Blot, Control, Immunofluorescence, Staining, Confocal Microscopy, Incubation, Flow Cytometry, Recombinant, Immunoprecipitation, Two Tailed Test

Journal: PLoS ONE

Article Title: Ezrin interacts with the tumor suppressor CHL1 and promotes neuronal differentiation of human neuroblastoma

doi: 10.1371/journal.pone.0244069

Figure Lengend Snippet: Using the SEQC patients’ datasets in the R2 Genomics Analysis and Visualization Platform ( http://r2.amc.nl ), event-free survival (top) and overall survival (bottom) curves were generated for (A) EZRIN and (B) MAP2 (Microtubule-associated protein-2) expression in all NB stages patients. Patients’ numbers (n) are shown in parentheses.

Article Snippet: To knock-down ezrin expression, cells were transfected with the silencing Ezrin shRNA lentiviral plasmid (shEzr) (a pool of three target-specific lentiviral vector plasmids by Santa Cruz Biotechnology, Dallas, TX, USA) or with the non-silencing control shRNA plasmid A (shNS) (Santa Cruz Biotechnology).

Techniques: Generated, Expressing

Journal: PLoS ONE

Article Title: Ezrin interacts with the tumor suppressor CHL1 and promotes neuronal differentiation of human neuroblastoma

doi: 10.1371/journal.pone.0244069

Figure Lengend Snippet: Immunofluorescence analysis of CHL1 and ezrin plasma membrane localization on (A) HTLA-230 and (B) IMR-32 cells using anti-CHL1 (green) (1) or anti-ezrin (red) (2) antibodies. HTLA-230 in photo A3 were double stained for CHL1 and ezrin (yellow, merge). IMR-32 in photo B3 expressed the transfected pCEFL-CHL1 molecule. Cells were counterstained with DAPI to visualize nuclei (blue) (Scale bars: 10 μm) (C) IMR-32 and HTLA-230 cells were lysed and subjected to Western blot (WB) analysis with anti-CHL1, anti-ezrin (Ezr), anti-radixin (Rdx), and anti-moesin (Moe) antibodies. Only HTLA-230 cell line displayed expression of endogenous CHL1 protein, whereas the three ERM proteins (Ezr, Rdx, and Moe) were equally expressed in both cell lines, and each of them was used as a loading control. (D) Cells lysates were subjected to each anti-ERM protein immunoprecipitation (IP) followed by anti-CHL1 Western blot or to anti-CHL1 immunoprecipitation followed by each anti-ERM protein Western blot. (E-F) Cell lysates from IMR-32 (as CHL1 negative control), and from HTLA-230 transfected with shNS (as non-silencing control) or shEzr, underwent Western blot analysis with anti-CHL1 and each anti-ERM protein antibody. HTLA-230 cells were lysed 7 days after transfection. 85–90% reduction in total ezrin levels was achieved compared to the non-silenced control, considered as 100%, whereas radixin and moesin levels did not change and they were used as loading controls. Data are representative of three independent experiments ± SD (*** p < 0.001). (G) Cell lysates from HTLA-230 transfected with shNS or shEzr were subjected to each anti-ERM protein immunoprecipitation followed by anti-CHL1 Western blot. All the immunoprecipitation results are representative of three independent experiments.

Article Snippet: To knock-down ezrin expression, cells were transfected with the silencing Ezrin shRNA lentiviral plasmid (shEzr) (a pool of three target-specific lentiviral vector plasmids by Santa Cruz Biotechnology, Dallas, TX, USA) or with the non-silencing control shRNA plasmid A (shNS) (Santa Cruz Biotechnology).

Techniques: Immunofluorescence, Clinical Proteomics, Membrane, Staining, Transfection, Western Blot, Expressing, Control, Immunoprecipitation, Negative Control

Journal: PLoS ONE

Article Title: Ezrin interacts with the tumor suppressor CHL1 and promotes neuronal differentiation of human neuroblastoma

doi: 10.1371/journal.pone.0244069

Figure Lengend Snippet: (A-C) Proteins from HTLA-230 cells transfected with shNS (control) or shEzr and from IMR-32 cells transfected with the empty vector pCEFL (control) or cotransfected with pCEFL-CHL1 and shNS or shEzr, were lysed 7 days after transfection, subjected to Western blot, and probed with anti-CHL1, anti-ezrin, anti-MAP2, anti-p53, anti-phospho-p38 (P-p38), and anti-phospho-ERK 1/2 (P-ERK 1/2) antibodies. (D) 75–80% ezrin expression reduction was achieved in shEzr-IMR-32 cells compared to the non-silenced controls (pCEFL was considered as 100%) with actin used as the loading control. (B-E) Protein levels were quantified by densitometry, normalized to the control values (fold induction = 1) and to the contents of the loading control proteins (actin or total p38 and ERK 1/2), and then visualized by histograms. Data are representative of three independent experiments ± SD. (*** p < 0.001).

Article Snippet: To knock-down ezrin expression, cells were transfected with the silencing Ezrin shRNA lentiviral plasmid (shEzr) (a pool of three target-specific lentiviral vector plasmids by Santa Cruz Biotechnology, Dallas, TX, USA) or with the non-silencing control shRNA plasmid A (shNS) (Santa Cruz Biotechnology).

Techniques: Transfection, Control, Plasmid Preparation, Western Blot, Expressing

Journal: PLoS ONE

Article Title: Ezrin interacts with the tumor suppressor CHL1 and promotes neuronal differentiation of human neuroblastoma

doi: 10.1371/journal.pone.0244069

Figure Lengend Snippet: Morphological characteristics of cells plated 48 hours after transfection and photographed three days later. (A) HTLA-230 cells transfected with shNS (1–2) or shEzr (3–4): shEzr-HTLA-230 grew faster and lost the neurite-like extensions detectable in control cells (see arrows in photo 2). (D) IMR-32 cells transfected with the negative control pCEFL (1–2) or cotransfected with pCEFL-CHL1 and shNS (3–4) or shEzr (5–6): over-expression of CHL1 in IMR-32 cells strongly reduced growth rate and induced neural differentiation with protrusion of neurite-like extensions (see arrows in photo 4), whereas the co-silencing of ezrin clearly reduced CHL1 effects. (Magnification 4x and 40x). (B-E) Histograms report the percentages of cells extending neurite-like protrusions. Data are representative of three independent experiments ± SD (*** p <0.001). (C-F) HTLA-230 or IMR-32 cells were plated 48 hours after transfection or cotransfection and cultured for 4 days. Every day, cells were trypsinized and counted. Data are representative of three independent experiments ± S.D. (* p < 0.05; *** p < 0.001).

Article Snippet: To knock-down ezrin expression, cells were transfected with the silencing Ezrin shRNA lentiviral plasmid (shEzr) (a pool of three target-specific lentiviral vector plasmids by Santa Cruz Biotechnology, Dallas, TX, USA) or with the non-silencing control shRNA plasmid A (shNS) (Santa Cruz Biotechnology).

Techniques: Transfection, Control, Negative Control, Over Expression, Cotransfection, Cell Culture

Journal: PLoS ONE

Article Title: Ezrin interacts with the tumor suppressor CHL1 and promotes neuronal differentiation of human neuroblastoma

doi: 10.1371/journal.pone.0244069

Figure Lengend Snippet: A migration assay was conducted for (A) HTLA-230 cells transfected with shNS or shEzr and for (B) IMR-32 cells transfected with pCEFL or cotransfected with pCEFL-CHL1 and shNS or shEzr. Cells were seeded 48 hours after transfection in the upper chamber of a Transwell insert: the ones which migrated to the lower side 24 hours later were detached and counted. (Three independent experiments ± S.D). (C) Colonies formed by HTLA-230 cells transfected with shNS or shEzr and by (D) IMR-32 cells transfected with pCEFL or cotransfected with pCEFL-CHL1 and shNS or shEzr. Cells were collected 48 hours after transfection and cultured in soft agar for 10 days. (Magnification 4x). Histograms show the mean colony number from three independent experiments ± S.D. (*** p < 0.001).

Article Snippet: To knock-down ezrin expression, cells were transfected with the silencing Ezrin shRNA lentiviral plasmid (shEzr) (a pool of three target-specific lentiviral vector plasmids by Santa Cruz Biotechnology, Dallas, TX, USA) or with the non-silencing control shRNA plasmid A (shNS) (Santa Cruz Biotechnology).

Techniques: Migration, Transfection, Cell Culture

Journal: PLoS ONE

Article Title: Ezrin interacts with the tumor suppressor CHL1 and promotes neuronal differentiation of human neuroblastoma

doi: 10.1371/journal.pone.0244069

Figure Lengend Snippet: The intra-cytoplasmic CHL1 C-terminal domain interacts with the N-terminal domain of ezrin, which links cytoskeletal actin through its C-terminal domain. This molecular binding activates MAP2 and p53 pathways and inhibits MAPK phosphorylation, inducing neuronal differentiation rather than cell proliferation and migration, thus conferring less aggressive features on NB (The CHL1 extracellular portion contains six Immunoglobulin (Ig)-like domains and five Fibronectin (FN) type III domains).

Article Snippet: To knock-down ezrin expression, cells were transfected with the silencing Ezrin shRNA lentiviral plasmid (shEzr) (a pool of three target-specific lentiviral vector plasmids by Santa Cruz Biotechnology, Dallas, TX, USA) or with the non-silencing control shRNA plasmid A (shNS) (Santa Cruz Biotechnology).

Techniques: Binding Assay, Phospho-proteomics, Migration