Journal: Nature cell biology

Article Title: Phosphatidylinositol 3-kinase Signaling is Spatially Organized at Endosomal Compartments by Microtubule-associated Protein 4

doi: 10.1038/s41556-020-00596-4

Figure Lengend Snippet: a, MAP4 loss impairs activation of Akt downstream of EGFR. Three different siRNAs were used individually to knockdown MAP4 in MDA-MB-231 cells. EGF induced Akt activation was analyzed by immunofluorescence microscopy. Scale bar, 5 μm; Error bars denote mean±SD; n=30 cells from representative experiments.

b, c, MAP4 loss impairs Akt activation downstream of integrin receptors. MDA-MB-231 cells were detached from culture plates 72-hours post-siRNA transfection for MAP4 knockdown (siMAP4#1) and resuspended in serum-free medium. Cells were seeded into culture plates-coated with type I collagen before harvesting at different time points followed by immunoblotting or immunofluorescence microscopy using an antibody specific for activated Akt and tubulin. Scale bar: 5 μm. Error bars denote mean±SD; n=4 independent experiments (b), n=30 cells from representative experiments (c)

d, MAP4 overexpression promotes Akt signaling. MDA-MB-231 cells ectopically overexpressing MAP4 or Mock were serumstarved overnight before stimulating with EGF. Cells were harvested at different time points and activation levels of Akt were examined by immunoblotting using activated Akt specific antibody. Error bars denote mean±SD; n=3 independent experiments

e, Effect of MAP4 knockdown in cell proliferation. MDA-MB- 468 and Cal51, both showing higher activation levels of Akt were transfected with siRNA for MAP4 knockdown. 72–96 hours postsiRNA transfection, cell numbers were manually quantified. Scale bar, 100 μm; Error bars denote mean±SD; n=3 independent experiments

f. g, h, Effect of MAP4 knockdown in cell invasion and cell migration. 48–72 hours post-siRNA transfection for MAP4 knockdown, cell invasion and scratch-wound healing for cell migration were performed. The image shown is the representative images of multiple reproducible experiments. Scale bar, 100 μm; Error bars denote mean±SD; n=9 fields from representative experiments (g), n=15 fields from representative experiments (h) Unprocessed_Western_Blots_Extended_Data_Fig8; Statistical_Source_Data_Extended_Data_Fig8

Article Snippet: Cloning of Human MAP4 and its Constructs Myc-DDK-tagged human microtubule-associated protein 4 (MAP4), the longest transcript variant 1 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_002375","term_id":"1677500572","term_text":"NM_002375"}} NM_002375 ) was synthesized and cloned into pCMV6-Entry vector by OriGene (Catalog No. RC216364).

Techniques: Knockdown, Activation Assay, Immunofluorescence, Microscopy, Transfection, Western Blot, Over Expression, Migration

Journal: Nature cell biology

Article Title: Phosphatidylinositol 3-kinase Signaling is Spatially Organized at Endosomal Compartments by Microtubule-associated Protein 4

doi: 10.1038/s41556-020-00596-4

Figure Lengend Snippet: a, b, Effect of MAP4 knockdown in endosomal localization of PI3Kα vesicles. siRNAs targeting the 3’ UTR region of MAP4 (siMAP4#3) were used to knockdown endogenous MAP4 in MDA-MB-231 cells expressing WT or the MTBD deletion mutant of MAP4. 48–72 hours post-transfection, cells were processed for immunofluorescence study using antibodies specific for endogenous p110α and clathrin or TFR. The quantification of the colocalization of p110α and CHC/TFR by Pearson’s r was shown in Figure 5c. The images shown are the representative images of multiple reproducible experiments. Scale bar, 5 μm

Article Snippet: Cloning of Human MAP4 and its Constructs Myc-DDK-tagged human microtubule-associated protein 4 (MAP4), the longest transcript variant 1 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_002375","term_id":"1677500572","term_text":"NM_002375"}} NM_002375 ) was synthesized and cloned into pCMV6-Entry vector by OriGene (Catalog No. RC216364).

Techniques: Knockdown, Expressing, Mutagenesis, Transfection, Immunofluorescence

Journal: Nature cell biology

Article Title: Phosphatidylinositol 3-kinase Signaling is Spatially Organized at Endosomal Compartments by Microtubule-associated Protein 4

doi: 10.1038/s41556-020-00596-4

Figure Lengend Snippet: a, Overexpression of MTBD impairs MAP4 association with PI3Kα. MAP4 and HA-tagged p110α were co-transfected along with empty GFP vector or GFP-MTBD or GFP-C-tail into Cos-7 cells. 24–48 hrs post-transfection, p110α were immunoprecipitated and co-immunoprecipitated MAP4 examined by immunoblotting. Error bars denote mean±SD; n=4 independent experiments

Article Snippet: Cloning of Human MAP4 and its Constructs Myc-DDK-tagged human microtubule-associated protein 4 (MAP4), the longest transcript variant 1 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_002375","term_id":"1677500572","term_text":"NM_002375"}} NM_002375 ) was synthesized and cloned into pCMV6-Entry vector by OriGene (Catalog No. RC216364).

Techniques: Over Expression, Transfection, Plasmid Preparation, Immunoprecipitation, Western Blot

Journal: Nature cell biology

Article Title: Phosphatidylinositol 3-kinase Signaling is Spatially Organized at Endosomal Compartments by Microtubule-associated Protein 4

doi: 10.1038/s41556-020-00596-4

Figure Lengend Snippet: a, in vivo association of PI3Kα with MAP4. Antibodies specific to p110α or p110β or p85α were used to immunoprecipitate wild type (MDA-MB-231) or mutant forms of PI3Kα (T47D and Cal51) and co-immunoprecipitated MAP4 examined by immunoblotting. The immunoblot shown is the representative of multiple experiments.

Article Snippet: Cloning of Human MAP4 and its Constructs Myc-DDK-tagged human microtubule-associated protein 4 (MAP4), the longest transcript variant 1 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_002375","term_id":"1677500572","term_text":"NM_002375"}} NM_002375 ) was synthesized and cloned into pCMV6-Entry vector by OriGene (Catalog No. RC216364).

Techniques: In Vivo, Mutagenesis, Immunoprecipitation, Western Blot

Journal: Nature cell biology

Article Title: Phosphatidylinositol 3-kinase Signaling is Spatially Organized at Endosomal Compartments by Microtubule-associated Protein 4

doi: 10.1038/s41556-020-00596-4

Figure Lengend Snippet: a, MAP4 loss affects PI3Kα vesicle distribution along microtubules. Three different individual siRNAs were used for MAP4 knockdown in MDA-MB-231 cells followed by immunofluorescence staining. The co-localization of p110α vesicles and tubulin were quantified by Pearson’s r. Scale bar, 5 μm; Error bars denote mean±SD; n=30 cells from representative experiments

Article Snippet: Cloning of Human MAP4 and its Constructs Myc-DDK-tagged human microtubule-associated protein 4 (MAP4), the longest transcript variant 1 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_002375","term_id":"1677500572","term_text":"NM_002375"}} NM_002375 ) was synthesized and cloned into pCMV6-Entry vector by OriGene (Catalog No. RC216364).

Techniques: Knockdown, Immunofluorescence, Staining

Journal: Nature cell biology

Article Title: Phosphatidylinositol 3-kinase Signaling is Spatially Organized at Endosomal Compartments by Microtubule-associated Protein 4

doi: 10.1038/s41556-020-00596-4

Figure Lengend Snippet: a, b, PLA shows induced association of MAP4 and IQGAP1 and that they are localized with EGFR. MDA-MB-231 cells were stimulated with EGF and processed for MAP4-IQGAP1 PLA followed by immunostaining with EGFR. The image shown is the representative images of multiple reproducible experiments. Scale bar, 5 μm; Error bars denote mean±SD; n=30 cells from representative experiments

c, d, e, MAP4 knockdown demonstrated by immunofluorescence study and immunoblotting. Three different siRNAs were used individually to knockdown MAP4 in MDA-MB-231 cells. 48–72 hours after siRNA transfection, cells were processed for immunofluorescence study using an antibody specific to MAP4 and tubulin. MAP4 knockdown was also shown by immunoblotting. The image and blot shown is the representative of reproducible experiments. Scale bar, 5 μm; Error bars denote mean±SD; n=30 cells from representative experiments

f, g, Localization of PI3Kα vesicles along microtubules in WT and MTBD deletion mutant of MAP4 expressing cells after knocking down endogenous MAP4. As described above, siRNAs targeting 3’ UTR region of MAP4 were used to knockdown endogenous MAP4. Cells were processed for immunofluorescence study using antibodies specific for p110α and tubulin. Distribution of PI3Kα vesicles along microtubules was quantified. n=30 cells from representative experiments

Article Snippet: Cloning of Human MAP4 and its Constructs Myc-DDK-tagged human microtubule-associated protein 4 (MAP4), the longest transcript variant 1 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_002375","term_id":"1677500572","term_text":"NM_002375"}} NM_002375 ) was synthesized and cloned into pCMV6-Entry vector by OriGene (Catalog No. RC216364).

Techniques: Knockdown, Expressing, Mutagenesis, Immunostaining, Immunofluorescence, Western Blot, Transfection

Journal: Nature cell biology

Article Title: Phosphatidylinositol 3-kinase Signaling is Spatially Organized at Endosomal Compartments by Microtubule-associated Protein 4

doi: 10.1038/s41556-020-00596-4

Figure Lengend Snippet: a, b, Coomassie staining of proteins co-immunoprecipitated with PI3Kα antibody (anti-p85α and anti-p110α antibodies used together) showed a distinct band above 170 kDa. Mass-spectrometry analysis of the isolated protein band revealed it as microtubule-associated protein 4 (MAP4). The image shown is the representative images of reproducible experiments. Arrow in image indicates the band of interest for mass spectrometry analysis.

c, PI3Kα in small vesicles distribute along MAP4 that mimics microtubules in different cell types. Immunofluorescence study was performed in different cell types using mouse anti-MAP4 and rabbit anti-p110α or p85α antibodies. The image shown is the representative images of reproducible experiments. Scale bar, 5 μm

d, Amino acid alignment of MAP4 MTBD along with that of Tau and MAP2. All four MTBD repeats of MAP4 (MTBD I- MTBD IV) and that of Tau and MAP2 (other microtubule-associated proteins expressed in neuronal cells) show highly similar amino acid order and microtubule-binding motif.

e, Representative MST binding affinity graphs for interaction between His-PI3Kα and GST-MAP4 proteins. Unprocessed_Gel_Image_Extended_Data_Fig5

Article Snippet: Cloning of Human MAP4 and its Constructs Myc-DDK-tagged human microtubule-associated protein 4 (MAP4), the longest transcript variant 1 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_002375","term_id":"1677500572","term_text":"NM_002375"}} NM_002375 ) was synthesized and cloned into pCMV6-Entry vector by OriGene (Catalog No. RC216364).

Techniques: Staining, Immunoprecipitation, Mass Spectrometry, Isolation, Immunofluorescence, Binding Assay

Journal: Nature cell biology

Article Title: Phosphatidylinositol 3-kinase Signaling is Spatially Organized at Endosomal Compartments by Microtubule-associated Protein 4

doi: 10.1038/s41556-020-00596-4

Figure Lengend Snippet: a, Schematic diagram of MAP4 protein showing N-terminal projection domain and C-terminal part that contains proline-rich (PR) region, MTBD and C-tail.

Article Snippet: Cloning of Human MAP4 and its Constructs Myc-DDK-tagged human microtubule-associated protein 4 (MAP4), the longest transcript variant 1 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_002375","term_id":"1677500572","term_text":"NM_002375"}} NM_002375 ) was synthesized and cloned into pCMV6-Entry vector by OriGene (Catalog No. RC216364).

Techniques:

Journal: Nature cell biology

Article Title: Phosphatidylinositol 3-kinase Signaling is Spatially Organized at Endosomal Compartments by Microtubule-associated Protein 4

doi: 10.1038/s41556-020-00596-4

Figure Lengend Snippet: a, b, MAP4 knockdown impairs EGF stimulated PI3,4,5P3 generation. Three different siRNAs were used individually to knockdown MAP4 in MDA-MB-231 cells. EGF induced PI3,4,5P3 was analyzed by immunofluorescence microscopy and MAP4 knockdown by immunoblotting. Scale bar, 5 μm; Error bars denote mean±SD; n=30 cells from representative experiments

Article Snippet: Cloning of Human MAP4 and its Constructs Myc-DDK-tagged human microtubule-associated protein 4 (MAP4), the longest transcript variant 1 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_002375","term_id":"1677500572","term_text":"NM_002375"}} NM_002375 ) was synthesized and cloned into pCMV6-Entry vector by OriGene (Catalog No. RC216364).

Techniques: Knockdown, Immunofluorescence, Microscopy, Western Blot

Journal: Nature cell biology

Article Title: Phosphatidylinositol 3-kinase Signaling is Spatially Organized at Endosomal Compartments by Microtubule-associated Protein 4

doi: 10.1038/s41556-020-00596-4

Figure Lengend Snippet: a, MAP4 loss impairs activation of Akt downstream of EGFR. Three different siRNAs individually used to knockdown MAP4 in MDA-MB-231 cells followed by examination of EGF induced Akt activation by immunoblotting. Error bars denote mean±SD; n=3 independent experiments

Article Snippet: Cloning of Human MAP4 and its Constructs Myc-DDK-tagged human microtubule-associated protein 4 (MAP4), the longest transcript variant 1 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_002375","term_id":"1677500572","term_text":"NM_002375"}} NM_002375 ) was synthesized and cloned into pCMV6-Entry vector by OriGene (Catalog No. RC216364).

Techniques: Activation Assay, Knockdown, Western Blot

Journal: bioRxiv

Article Title: TRiPPing the sensors: The osmosensing pathway of Polycystin 2

doi: 10.1101/2023.05.09.540007

Figure Lengend Snippet: A. MAP4 binds and stabilizes microtubules. Disassociation of MAP4 from the microtubules due to phosphorylation of MAP4 destabilizes microtubules. B. Expression of p-MAP4 in C2C12 CTL and PC2 KO cells at 300 mOsm and 400 mOsm. Total protein was used as loading control. C. p-MAP4 increased in C2C12 CTL cells after increasing extracellular osmolarity. Basal p-MAP4 levels were higher in PC2 KO cells compared to CTL but did not change with osmolarity. Bars represent mean±SEM. Data were analyzed to determine normality. Statistical analysis was determined by Two-way ANOVA followed by Sidak’s test. p-values listed in the figure. D. Western blot of MAP4 expression in C2C12 CTL and PC2 KO cells at 300 mOsm and 400 mOsm. Total protein was used as loading control. E. MAP4 expression was significantly decreased in PC2 KO cells. Bars represent mean±SEM. Data were analyzed to determine normality. Statistical analysis was determined by Two-way ANOVA followed by Sidak’s test. p-values listed in the figure. F. Expression of CLIMP63 remained unchanged in C2C12 CTL and PC2 KO cells. Bars represent mean±SEM. Data were analyzed to determine normality. Statistical analysis was determined by Two-way ANOVA followed by Sidak’s test.

Article Snippet: The cells were incubated with antibodies against PC2 YCE2 (1:100, sc-47734, Santa Cruz Biotechnology), MAP4 (1:100; #SC-390286; Santa Cruz), CLIMP63 (CKAP4, 1:2,000, A302-257A; Fortis Life Science), AQP2 (1,1000, #3487; CST), Arl13b (1:100, #17711-1-AP; ProteinTech) overnight at 4°C, followed by the appropriate secondary antibody (Alexa Fluor 488 donkey anti-mouse IgG (1:1000, A21202, Invitrogen), Alexa Fluor 546 donkey anti-rabbit IgG (1:800, A10040, Invitrogen), for 1 hr, then washed three times in PBS.

Techniques: Phospho-proteomics, Expressing, Control, Western Blot

Journal: bioRxiv

Article Title: TRiPPing the sensors: The osmosensing pathway of Polycystin 2

doi: 10.1101/2023.05.09.540007

Figure Lengend Snippet: A. Expression of MAP4 in C2C12 CTL and MAP4-KO cells. GAPDH was used as loading control. B. MAP4 expression was decreased 75% in the knock down cell line. C. Immunofluorescence staining of MAP4 and CLIMP63 in C2C12 CTL cells (top panels) and MAP4-KO cells (bottom panels). D. Representative trace of cytosolic calcium changes in C2C12 MAP4-KO cells ( blue line ). Scale bars represent 10 μm. E. Area under the curve was significantly decreased in the MAP4-KO after increasing extracellular osmolarity. Bars represent mean±SEM. Data were analyzed to determine normality. Statistical analysis was determined by student’s t-test. p-values listed in figure. F. Representative trace of cytosolic calcium increase using thapsigargin after increasing extracellular osmolarity in MAP4-KO cells. G. Representative images of EB3-tdTomato expression in C2C12 MAP4-KO cells at 300 mOsm (left panel) and 400 mOsm (right panels). Scale bars represent 10 μm. H. EB3 comet length was significantly decreased in C2C12 MAP4-KO at basal osmolarity (300 mOsm) and remained further unchanged after osmotic increase. Bars represent mean±SEM. Data were analyzed to determine normality. Statistical analysis was determined by One-way ANOVA. p-values listed in figure.

Article Snippet: The cells were incubated with antibodies against PC2 YCE2 (1:100, sc-47734, Santa Cruz Biotechnology), MAP4 (1:100; #SC-390286; Santa Cruz), CLIMP63 (CKAP4, 1:2,000, A302-257A; Fortis Life Science), AQP2 (1,1000, #3487; CST), Arl13b (1:100, #17711-1-AP; ProteinTech) overnight at 4°C, followed by the appropriate secondary antibody (Alexa Fluor 488 donkey anti-mouse IgG (1:1000, A21202, Invitrogen), Alexa Fluor 546 donkey anti-rabbit IgG (1:800, A10040, Invitrogen), for 1 hr, then washed three times in PBS.

Techniques: Expressing, Control, Knockdown, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: TRiPPing the sensors: The osmosensing pathway of Polycystin 2

doi: 10.1101/2023.05.09.540007

Figure Lengend Snippet: A. Representative immunofluorescent staining of imCD3 CTL cells at basal osmotic conditions (top panels) labeling AQP2 (red) and MAP4 (green) and 10 min after increasing extracellular osmolarity to 400 mOsm (bottom panels). Scale bars represent 10 μm. B. Immunofluorescent staining of A (red) and MAP4 (green) in PC2 KO imCD3 cells at basal osmotic conditions (top panels) and after increase of extracellular osmolarity (bottom panels). Scale bars represent 10 μm. C. Quantification of AQP2 vesicle size significantly increases in imCD3 CTL cells after increasing extracellular osmolarity (400 mOsm) for 10 min. Vesicle size remained unchanged in PC2 KO cells. Bars represent mean±SEM. Data were analyzed to determine normality and statistical analysis was determined by 2-way ANOVA. p-values listed in the figure. D. Immunofluorescent staining of AQP2 insertion in the presence of 100nM AVP of PC2 KO imCD3 cells. Scale bars represent 10 μm. E. Immunofluorescent staining of AQP2 (red) and MAP4 (green) in isolated renal tubules from CTL mice. AQP2 and MAP4 colocalized into the membrane of the collecting duct cell membrane (top panels). Scale bars represent 10 μm. F. Immunofluorescent staining of AQP2 (red) and MAP4 (green) in isolated renal tubules from tubule specific PC2 KO mice. AQP2 and MAP4 have diffused staining in the cytosol of collecting duct cells (top panels). Scale bars represent 10 μm.

Article Snippet: The cells were incubated with antibodies against PC2 YCE2 (1:100, sc-47734, Santa Cruz Biotechnology), MAP4 (1:100; #SC-390286; Santa Cruz), CLIMP63 (CKAP4, 1:2,000, A302-257A; Fortis Life Science), AQP2 (1,1000, #3487; CST), Arl13b (1:100, #17711-1-AP; ProteinTech) overnight at 4°C, followed by the appropriate secondary antibody (Alexa Fluor 488 donkey anti-mouse IgG (1:1000, A21202, Invitrogen), Alexa Fluor 546 donkey anti-rabbit IgG (1:800, A10040, Invitrogen), for 1 hr, then washed three times in PBS.

Techniques: Staining, Labeling, Isolation, Membrane

Journal: bioRxiv

Article Title: TRiPPing the sensors: The osmosensing pathway of Polycystin 2

doi: 10.1101/2023.05.09.540007

Figure Lengend Snippet: A. Expression of PC2 in C2C12 CTL and MAP4-KO cells. Tubulin was used as loading control. B. PC2 expression was significantly increased in MAP4-KO cells in comparison to CTL cells. Bars represent mean±SEM. Data were analyzed to determine normality. Statistical analysis was determined by student’s t-test. p-values listed in figure. C. Immunofluorescent staining of PC2 (green) and CLIMP63 (ER; red) in C2C12 CTL cells (top panels) and MAP4-KO cells (bottom panels). Scale bars represent 10 μm.

Article Snippet: The cells were incubated with antibodies against PC2 YCE2 (1:100, sc-47734, Santa Cruz Biotechnology), MAP4 (1:100; #SC-390286; Santa Cruz), CLIMP63 (CKAP4, 1:2,000, A302-257A; Fortis Life Science), AQP2 (1,1000, #3487; CST), Arl13b (1:100, #17711-1-AP; ProteinTech) overnight at 4°C, followed by the appropriate secondary antibody (Alexa Fluor 488 donkey anti-mouse IgG (1:1000, A21202, Invitrogen), Alexa Fluor 546 donkey anti-rabbit IgG (1:800, A10040, Invitrogen), for 1 hr, then washed three times in PBS.

Techniques: Expressing, Control, Comparison, Staining

Journal: bioRxiv

Article Title: MAP4-MAP7D1 partitioning on tyrosinated-detyrosinated microtubules coordinates lysosome positioning in nutrient signalling

doi: 10.1101/2025.10.07.680844

Figure Lengend Snippet: MAP4 projection domains restrict its association to tyrosinated microtubules. (a) The Schematic of MAP4 truncation constructs used in the study. (b-e) Representative confocal images of BS-C-1 cells transiently expressing GFP-tagged MAP4 constructs (magenta) immunostained for detyrosinated microtubules (green) along with corresponding insets show magnified views of the regions indicated by yellow boxes. (f) Box plot quantifying MAP4 colocalization with detyrosinated microtubules. (g) Model illustrating the role of MAP4 projection domains in conferring specificity for tyrosinated microtubules. Data represents mean (line) ± SD (box). Number of cells (n) analyzed for MAP4-FL (10), MAP4-MTBD (11), MAP4-ΔN (15) and MAP4-ΔC (14). Statistical significance was assessed using the Mann– Whitney U test (*p<0.05, **p<0.01, not significant [ns]). Scale bars: 10 μm.

Article Snippet: Microtubule Binding Domain (MTBD) of MAP4 mRuby-MAP4-C-10 (Addgene Plasmid #55873 from Michael Davidson).

Techniques: Construct, Expressing, MANN-WHITNEY

Journal: bioRxiv

Article Title: MAP4-MAP7D1 partitioning on tyrosinated-detyrosinated microtubules coordinates lysosome positioning in nutrient signalling

doi: 10.1101/2025.10.07.680844

Figure Lengend Snippet: Kinesin-1 and kinesin-3 rigor mutants differentially associate with MAP4 and MAP7D1 (a-b) Confocal images of BS-C-1 cells transiently expressing GFP-tagged (a) KIF5B-R (green) or (b) KIF1A-R (green), immunostained for endogenous MAP4 (magenta). (c) Bar plot of Mander’s colocalization quantification shows significantly higher colocalization of MAP4 with KIF1A-R ( n = 8 cells) than KIF5B-R ( n = 9 cells). (d-e) Confocal images of cells expressing GFP-tagged (d) KIF1A-R or (e) KIF5B-R (green), immunostained for endogenous MAP7D1 (magenta). (f) Bar plot of Mander’s colocalization quantification reveals significantly higher colocalization of MAP7D1 with KIF5B-R ( n = 10 cells) than KIF1A-R ( n = 9 cells). (g–h) Co-expression of KIF5B-R (green) with (g) MAP4-FL or (h) MAP4-MTBD (magenta), Line intensity profiles show MAP4-FL is excluded from KIF5B-R-decorated microtubules, while MAP4-MTBD exhibits extensive colocalization. (i) Model illustrating the preferential association of KIFB-R and KIF1A-R with MAP7D1 and MAP4, respectively. Bars represent the mean; whiskers indicate standard deviation from three independent experiments. Statistical significance was assessed using the Mann–Whitney U test (****p < 0.0001). Scale bars: 10 μm.

Article Snippet: Microtubule Binding Domain (MTBD) of MAP4 mRuby-MAP4-C-10 (Addgene Plasmid #55873 from Michael Davidson).

Techniques: Expressing, Standard Deviation, MANN-WHITNEY