Review

human map4 (OriGene)

OriGene is a verified supplier

OriGene manufactures this product

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

OriGene human map4

a, <t>MAP4</t> 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. </p/>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) </p/>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 </p/>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 </p/>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

Human Map4, supplied by OriGene, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/map4/Mtap4+(BC044654)+Mouse+Tagged+ORF+Clone/pmc08647654-510-2-30
Average 90 stars, based on 4 article reviews

human map4 - by Bioz Stars, 2026-11

90/100 stars

Images

1) Product Images from "Phosphatidylinositol 3-kinase Signaling is Spatially Organized at Endosomal Compartments by Microtubule-associated Protein 4"

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

Journal: Nature cell biology

doi: 10.1038/s41556-020-00596-4

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. </p/>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) </p/>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 </p/>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 </p/>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

Figure Legend 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

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

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

Figure Legend 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

Techniques Used: Knockdown, Expressing, Mutagenesis, Transfection, Immunofluorescence

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

Figure Legend 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

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

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.

Figure Legend 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.

Techniques Used: In Vivo, Mutagenesis, Immunoprecipitation, Western Blot

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

Figure Legend 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

Techniques Used: Knockdown, Immunofluorescence, Staining

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 </p/>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 </p/>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

Figure Legend 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

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

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

Figure Legend 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

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

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.

Figure Legend 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.

Techniques Used:

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

Figure Legend 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

Techniques Used: Knockdown, Immunofluorescence, Microscopy, Western Blot

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

Figure Legend 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

Techniques Used: Activation Assay, Knockdown, Western Blot

Cloning:

Article Title: Phosphatidylinositol 3-kinase Signaling is Spatially Organized at Endosomal Compartments by Microtubule-associated Protein 4
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). .. The integrity of DNA sequence was validated by OriGene and us.

Construct:

Article Title: Mechanism of spindle pole organization and instability in human oocytes.
Article Snippet: INTRODUCTION: Many human eggs carry an incorrect number of chromosomes, a condition known as aneuploidy.. Aneuploidy in human eggs is the leading cause of aberrant embryonic development, resulting in pregnancy loss and genetic disorders such as Down syndrome.. Most aneuploidy results from chromosome segregation errors during the maturation of oocytes into fertilizable eggs.

Variant Assay:

Synthesized:

Clone Assay:

Expressing:

Article Title: Compartmentalized control of Cdk1 drives mitotic spindle assembly.
Article Snippet: Mild inhibition of Cdk1 activity was obtained by adding RO-3306 at 0.5 mM (low-RO; Calbiochem). .. Mammalian expression plasmids for turbo-GFP-Map4-wild type (tGFP-Map4-WT), turbo-GFP-Map4 rendered non-phosphorylatable at serine 787 (tGFP-Map4-S787A) and turbo-GFP-Map4 defective in PP1 binding Map4 (tGFP-Map4-PP1-KO), in which the Map4 peptide KDVRW (from aa 318 to 322; human Map4 isoform 1) was mutated into KDARA, were generated and purchased from OriGene. .. Cdk1WT and cdk1AF mammalian expression plasmids have been described before (Hagting et al., 1998) and were purchased from Addgene.

Binding Assay:

Generated:

Recombinant:

Purification:

other:

Article Title: Compartmentalized control of Cdk1 drives mitotic spindle assembly.
Article Snippet: Cdk1WT and cdk1AF mammalian expression plasmids have been described before (Hagting et al., 1998) and were purchased from Addgene.

Article Title: Compartmentalized control of Cdk1 drives mitotic spindle assembly.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Software and algorithms ImageJ ImageJ; https://imagej.nih.gov ZEN 3.1 Zeiss https://www.zeiss.com BioRender BioRender https://biorender.com Excel Microsoft https://microsoft.com

Image Search Results

MAP7D1 and MAP4 nanoclusters are largely exclusive on microtubule network. (a) Dual-colour confocal images of BS-C-1 cells showing the distribution of microtubule-bound MAP7D1 (magenta) and MAP4 (green). Magenta arrows indicating MAP7D1-dominated microtubules, white arrows indicating MAP4-dominated microtubules, and yellow arrowheads marking microtubules decorated by both MAPs. A corresponding line intensity plot along the yellow line in the zoomed image reveals that the two MAPs are predominantly associated with distinct microtubule tracks. (b) Two-colour STORM image showing the nanoscale distribution of MAP7D1 (magenta) and MAP4 (green) along microtubule tracks. The two MAPs form discrete nanoclusters along microtubule tracks. (c–e) Representative dual-colour STORM images of individual microtubule segments showing examples of MAP7D1-dominant (c), MAP4-dominant (d), and regions with both MAPs (e). (f) Pie-chart depicting the distribution of microtubule segments populated with either MAP4 or MAP7D1 or both. (g) The percentage localization density of each MAP was quantified for individual ROIs and plotted as a stacked column graph. ROIs were sorted in ascending order of MAP4 localization density, revealing an inverse correlation between MAP4 and MAP7D1 localization distribution along microtubules. (h) Representative two-colour STORM images of MAP4 and MAP7D1 nanoclusters classified into four categories based on the extent of spatial overlap. Corresponding histograms show the percentage overlap between MAP4 and MAP7D1 localizations plotted along the x-axis. (i) Pie chart shows the proportion of MAP4–MAP7D1 nanocluster overlap: non-overlapping, small overlap (<20%), partial overlap (20–80%), and high or complete overlap (>80%). This data further supports the largely non-overlapping nature of the two MAPs. Scale bars: 10 μm for (a), 5 μm for zoomed images; 1 μm for (b); 500 nm for (c–e); 100 nm for (g).

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: MAP7D1 and MAP4 nanoclusters are largely exclusive on microtubule network. (a) Dual-colour confocal images of BS-C-1 cells showing the distribution of microtubule-bound MAP7D1 (magenta) and MAP4 (green). Magenta arrows indicating MAP7D1-dominated microtubules, white arrows indicating MAP4-dominated microtubules, and yellow arrowheads marking microtubules decorated by both MAPs. A corresponding line intensity plot along the yellow line in the zoomed image reveals that the two MAPs are predominantly associated with distinct microtubule tracks. (b) Two-colour STORM image showing the nanoscale distribution of MAP7D1 (magenta) and MAP4 (green) along microtubule tracks. The two MAPs form discrete nanoclusters along microtubule tracks. (c–e) Representative dual-colour STORM images of individual microtubule segments showing examples of MAP7D1-dominant (c), MAP4-dominant (d), and regions with both MAPs (e). (f) Pie-chart depicting the distribution of microtubule segments populated with either MAP4 or MAP7D1 or both. (g) The percentage localization density of each MAP was quantified for individual ROIs and plotted as a stacked column graph. ROIs were sorted in ascending order of MAP4 localization density, revealing an inverse correlation between MAP4 and MAP7D1 localization distribution along microtubules. (h) Representative two-colour STORM images of MAP4 and MAP7D1 nanoclusters classified into four categories based on the extent of spatial overlap. Corresponding histograms show the percentage overlap between MAP4 and MAP7D1 localizations plotted along the x-axis. (i) Pie chart shows the proportion of MAP4–MAP7D1 nanocluster overlap: non-overlapping, small overlap (<20%), partial overlap (20–80%), and high or complete overlap (>80%). This data further supports the largely non-overlapping nature of the two MAPs. Scale bars: 10 μm for (a), 5 μm for zoomed images; 1 μm for (b); 500 nm for (c–e); 100 nm for (g).

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

Techniques:

MAP4 preferentially associates with tyrosinated microtubules. Two-color super-resolution SIM images of endogenous MAP4 (magenta) with (a) tyrosinated, (b) detyrosinated, and (c) acetylated microtubules (green) in BS-C-1 cells, with corresponding line intensity plots showing preferential MAP4 localization to tyrosinated microtubules. (d) Three-color SIM image of MAP4 (magenta) with tyrosinated (green) and acetylated (blue) microtubules. Arrowheads indicate MAP4 associated with acetylated-tyrosinated microtubules; arrows indicate MAP4 associated with acetylated-detyrosinated microtubules. (e) Quantification of MAP4 localization across distinct microtubule subsets from n = 29 ROIs per subset from 12 different cells. MAP4 fluorescence intensity was measured from ROIs of equal size drawn on each microtubule population, ensuring uniform sampling across microtubule populations. (f) Percentage of MAP4 associated with each subset was calculated by dividing the mean MAP4 intensity on that subset by the total MAP4 intensity across all ROIs. (g) Two-color STORM images of MAP4 (magenta) with tyrosinated microtubules (green). HDBSCAN-based cluster analysis quantifies (h) inter-cluster distance and (i) cluster density (number of clusters per unit length), revealing significantly higher MAP4 clustering on tyrosinated microtubules (yellow box) compared to detyrosinated microtubules (cyan box). ROIs for detyrosinated microtubules were selected based on MAP4-decorated microtubule patterns lacking tyrosination signal. n = 26 ROIs were analyzed per microtubule subset from a cell. Data represents mean (line) ± SD (box). Statistical significance was assessed using the Mann–Whitney U test (****p < 0.0001). Scale bars: 10 μm for (a–c) and corresponding zoomed images at 5 μm; 5 μm for (d), 2 μm for (g), and corresponding zoomed images at 500 nm.

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 preferentially associates with tyrosinated microtubules. Two-color super-resolution SIM images of endogenous MAP4 (magenta) with (a) tyrosinated, (b) detyrosinated, and (c) acetylated microtubules (green) in BS-C-1 cells, with corresponding line intensity plots showing preferential MAP4 localization to tyrosinated microtubules. (d) Three-color SIM image of MAP4 (magenta) with tyrosinated (green) and acetylated (blue) microtubules. Arrowheads indicate MAP4 associated with acetylated-tyrosinated microtubules; arrows indicate MAP4 associated with acetylated-detyrosinated microtubules. (e) Quantification of MAP4 localization across distinct microtubule subsets from n = 29 ROIs per subset from 12 different cells. MAP4 fluorescence intensity was measured from ROIs of equal size drawn on each microtubule population, ensuring uniform sampling across microtubule populations. (f) Percentage of MAP4 associated with each subset was calculated by dividing the mean MAP4 intensity on that subset by the total MAP4 intensity across all ROIs. (g) Two-color STORM images of MAP4 (magenta) with tyrosinated microtubules (green). HDBSCAN-based cluster analysis quantifies (h) inter-cluster distance and (i) cluster density (number of clusters per unit length), revealing significantly higher MAP4 clustering on tyrosinated microtubules (yellow box) compared to detyrosinated microtubules (cyan box). ROIs for detyrosinated microtubules were selected based on MAP4-decorated microtubule patterns lacking tyrosination signal. n = 26 ROIs were analyzed per microtubule subset from a cell. Data represents mean (line) ± SD (box). Statistical significance was assessed using the Mann–Whitney U test (****p < 0.0001). Scale bars: 10 μm for (a–c) and corresponding zoomed images at 5 μm; 5 μm for (d), 2 μm for (g), and corresponding zoomed images at 500 nm.

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

Techniques: Fluorescence, Sampling, MANN-WHITNEY

MAP7D1, not MAP4, is sensitive to lattice expansion. (a-d) Three-color confocal images and corresponding line intensity profiles of BS-C-1 cells (a) DMSO-treated cells immunostained for endogenous MAP4 (magenta) with acetylated (green) and tyrosinated (blue) microtubules. Arrowheads indicate MAP4 colocalization with acetylated-tyrosinated microtubules; arrows indicate MAP4 exclusion from acetylated-detyrosinated microtubules. (b) Taxol-induced lattice expansion preserves MAP4’s preference for acetylated-tyrosinated microtubules. (c) DMSO-treated cells immunostained for endogenous MAPD1 (magenta) with acetylated (green) and tyrosinated (blue) microtubules. Arrowheads indicate MAP7D1 sparsely decorated on acetylated-tyrosinated microtubules; arrows indicate MAP7D1 enriched on acetylated-detyrosinated microtubules. (d) Taxol-induced lattice expansion redistributes MAP7D1 to acetylated-detyrosinated (arrows) and acetylated-tyrosinated (arrowheads) microtubules. (e) Super-resolution STORM images showing endogenous MAP4 (magenta) or MAP7D1 (magenta) localization on tyrosinated microtubules in DMSO or Taxol-treated cells. ROIs were drawn on tyrosinated microtubules based on corresponding two-color TIRF images (shown as insets, also see Figure S3b, c). Yellow box shows the corresponding zoomed ROIs, highlighting unaltered MAP4 localization but increased MAP7D1 density on tyrosinated microtubules after lattice expansion. (h) Quantification of MAP4 and MAP7D1 localization density on tyrosinated microtubules from STORM images. No. of ROIs analyzed, MAP4 DMSO ( n =383), MAP4 Taxol (n= 658), MAP7D1 DMSO (n= 317), MAP7D1 Taxol (n= 374) from 3 different cells. Data represents mean (line) ± SD (box). Statistical significance was assessed using the Mann–Whitney U test (****p < 0.0001, not significant [ns]). Scale bars: 10 μm for (a–d), with zoomed images at 2 μm; 2 μm for (e), with zoomed images at 500 nm.

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: MAP7D1, not MAP4, is sensitive to lattice expansion. (a-d) Three-color confocal images and corresponding line intensity profiles of BS-C-1 cells (a) DMSO-treated cells immunostained for endogenous MAP4 (magenta) with acetylated (green) and tyrosinated (blue) microtubules. Arrowheads indicate MAP4 colocalization with acetylated-tyrosinated microtubules; arrows indicate MAP4 exclusion from acetylated-detyrosinated microtubules. (b) Taxol-induced lattice expansion preserves MAP4’s preference for acetylated-tyrosinated microtubules. (c) DMSO-treated cells immunostained for endogenous MAPD1 (magenta) with acetylated (green) and tyrosinated (blue) microtubules. Arrowheads indicate MAP7D1 sparsely decorated on acetylated-tyrosinated microtubules; arrows indicate MAP7D1 enriched on acetylated-detyrosinated microtubules. (d) Taxol-induced lattice expansion redistributes MAP7D1 to acetylated-detyrosinated (arrows) and acetylated-tyrosinated (arrowheads) microtubules. (e) Super-resolution STORM images showing endogenous MAP4 (magenta) or MAP7D1 (magenta) localization on tyrosinated microtubules in DMSO or Taxol-treated cells. ROIs were drawn on tyrosinated microtubules based on corresponding two-color TIRF images (shown as insets, also see Figure S3b, c). Yellow box shows the corresponding zoomed ROIs, highlighting unaltered MAP4 localization but increased MAP7D1 density on tyrosinated microtubules after lattice expansion. (h) Quantification of MAP4 and MAP7D1 localization density on tyrosinated microtubules from STORM images. No. of ROIs analyzed, MAP4 DMSO ( n =383), MAP4 Taxol (n= 658), MAP7D1 DMSO (n= 317), MAP7D1 Taxol (n= 374) from 3 different cells. Data represents mean (line) ± SD (box). Statistical significance was assessed using the Mann–Whitney U test (****p < 0.0001, not significant [ns]). Scale bars: 10 μm for (a–d), with zoomed images at 2 μm; 2 μm for (e), with zoomed images at 500 nm.

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

Techniques: MANN-WHITNEY

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.

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

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.

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

(a) Correlative live-cell and STORM imaging of lysosomes with MAP4 in BS-C-1 cells. Single-particle trajectories (SPT) from live imaging were aligned and overlaid on the corresponding STORM image of MAP4. Anterograde track is shown in yellow and retrograde track in cyan. In corresponding graphs, the trajectories are colour-coded to indicate run phases (blue) versus pause phases (red), revealing more processive retrograde runs than anterograde runs. (b) Scatter plot of lysosome speed versus MAP4 density. Dotted lines mark the mean on both axes. Points below (pink) or above (blue) the mean MAP4 density illustrate that lysosomes show reduced average speed at higher MAP4 density. (c) Paired correlation plot between lysosomal speed and MAP4 density, separated into anterograde (n = 13 trajectories) and retrograde (n = 11 trajectories) tracks. This shows that anterograde-moving lysosomes experience greater hindrance in speed with increasing MAP4 density. (d) Paired correlation plot between run-phase and MAP4 density for the same anterograde (n = 13) and retrograde (n = 11) tracks. Higher MAP4 density reduces run-phase more markedly for anterograde than for retrograde tracks. (e) Correlative live-cell and STORM imaging of lysosomes with MAP7D1 (magenta) in BS-C-1 cells. Single-particle trajectories of lysosomes from live imaging were aligned and overlaid on the STORM image of MAP7D1. Anterograde trajectories are marked in yellow and retrograde in cyan. Run phases (blue) and pause phases (red) highlight that retrograde runs are more processive than anterograde runs. (f) Scatter plot of lysosome speed versus MAP7D1 density. Dotted lines mark the mean on both axes. Points below (pink) or above (blue) the mean MAP7D1 density indicate that lysosomes have reduced average speed at higher MAP7D1 density. (g) Paired correlation plot between lysosomal speed and MAP7D1 density, separated into anterograde (n = 14 trajectories) and retrograde (n = 17 trajectories) tracks. Anterograde-moving lysosomes show greater hindrance in speed as MAP7D1 density increases. (h) Paired correlation plot between run-phase and MAP7D1 density for the same anterograde (n = 14) and retrograde (n = 17) tracks. Higher MAP7D1 density reduces run-phase measures more for anterograde than retrograde tracks. Scale bars: 500 nm.

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: (a) Correlative live-cell and STORM imaging of lysosomes with MAP4 in BS-C-1 cells. Single-particle trajectories (SPT) from live imaging were aligned and overlaid on the corresponding STORM image of MAP4. Anterograde track is shown in yellow and retrograde track in cyan. In corresponding graphs, the trajectories are colour-coded to indicate run phases (blue) versus pause phases (red), revealing more processive retrograde runs than anterograde runs. (b) Scatter plot of lysosome speed versus MAP4 density. Dotted lines mark the mean on both axes. Points below (pink) or above (blue) the mean MAP4 density illustrate that lysosomes show reduced average speed at higher MAP4 density. (c) Paired correlation plot between lysosomal speed and MAP4 density, separated into anterograde (n = 13 trajectories) and retrograde (n = 11 trajectories) tracks. This shows that anterograde-moving lysosomes experience greater hindrance in speed with increasing MAP4 density. (d) Paired correlation plot between run-phase and MAP4 density for the same anterograde (n = 13) and retrograde (n = 11) tracks. Higher MAP4 density reduces run-phase more markedly for anterograde than for retrograde tracks. (e) Correlative live-cell and STORM imaging of lysosomes with MAP7D1 (magenta) in BS-C-1 cells. Single-particle trajectories of lysosomes from live imaging were aligned and overlaid on the STORM image of MAP7D1. Anterograde trajectories are marked in yellow and retrograde in cyan. Run phases (blue) and pause phases (red) highlight that retrograde runs are more processive than anterograde runs. (f) Scatter plot of lysosome speed versus MAP7D1 density. Dotted lines mark the mean on both axes. Points below (pink) or above (blue) the mean MAP7D1 density indicate that lysosomes have reduced average speed at higher MAP7D1 density. (g) Paired correlation plot between lysosomal speed and MAP7D1 density, separated into anterograde (n = 14 trajectories) and retrograde (n = 17 trajectories) tracks. Anterograde-moving lysosomes show greater hindrance in speed as MAP7D1 density increases. (h) Paired correlation plot between run-phase and MAP7D1 density for the same anterograde (n = 14) and retrograde (n = 17) tracks. Higher MAP7D1 density reduces run-phase measures more for anterograde than retrograde tracks. Scale bars: 500 nm.

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

Techniques: Imaging, Single Particle

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: MAP density on microtubules regulates nutrient-dependent lysosomal positioning (a) Representative confocal images of BS-C-1 cells immunostained for endogenous LAMP2 (green) showing lysosomal distribution and (b) radial intensity profile plot for quantification of lysosomal distribution under control (n=27 cells), nutrient starvation (n=17 cells) and 2x amino acids stimulated (n=17 cells) conditions, (c-f) Representative STORM images MAP4 and MAP7D1 under control, nutrient starvation and nutrient stimulation along with corresponding localization density distribution plots from 3 different cells in each conditions. (g-h) Representative confocal images of control cells showing lysosomal distribution along with the corresponding radial intensity profile plot in endogenous (n=27 cells), MAP4-FL overexpression (n=27 cells) and MAP7-FL overexpression (n=23 cells) conditions. (i-j) Representative confocal images of untreated control cells showing lysosomal distribution along with corresponding radial intensity profile plot in scramble siRNA (n=21 cells), MAP4 siRNA (n=37 cells) and MAP7D1 siRNA (n=27 cells). (k-l) Representative confocal images of nutrient-stimulated cells showing lysosomal distribution along with corresponding radial intensity profile plot in scramble siRNA (n=23 cells), MAP4 siRNA (n=41 cells) and MAP7D1 siRNA (n=31 cells). Lysosomal distribution as a function of distance from the center of the nucleus was quantified from confocal images using the Radial Profile plugin in ImageJ (see Methods for details). Radial intensity profile graphs represent the mean ± SEM of lysosomal intensity profiles, averaged from three independent experiments. Statistical significance was assessed using the Kolmogorov–Smirnov (KS) test, comparing treated conditions to the control (n.s.> 0.05, *** < 0.05, **** p < 0.00001). Scale bars: 10 μm for confocal images, 5 μm for STORM images in c, e and corresponding insets at 1 μm.

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

Techniques: Control, Over Expression

Proposed model The schematic presents a model integrating findings from this study and prior reports , , , . Under homeostatic conditions, the microtubule network is differentially decorated with MAPs: MAP4 is enriched on tyrosinated microtubules, while MAP7D1 is enriched on detyrosinated microtubules. These MAP–PTM combinations establish specialized tracks that spatiotemporally regulate lysosome transport. On MAP7D1-decorated detyrosinated microtubules, kinesin-1 is preferentially recruited but its motility is dampened by the high density of MAP7D1 nanoclusters, promoting lysosomal immobilization in the perinuclear region to support autophagy-related functions. In contrast, MAP4-enriched tyrosinated microtubules favor fast kinesin-3 or kinesin-2–driven anterograde transport, redistributing lysosomes toward the cell periphery to enhance mTORC1 signaling. Importantly, anterograde transport is dynamically balanced by dynein-driven retrograde movement, which counteracts kinesin activity and maintains equilibrium between perinuclear and peripheral lysosome pools during homeostasis. Upon nutrient modulation, MAP densities are remodelled along microtubule tracks. Nutrient deprivation increases MAP7D1 density and reduces MAP4 abundance, thereby immobilizing kinesin-1 and biasing transport toward dynein-driven retrograde motility, which retains lysosomes in the perinuclear region. Conversely, nutrient stimulation decreases MAP7D1 density, while maintaining optimal MAP4 density, thereby enabling faster motility of kinesin and promoting anterograde transport and peripheral positioning of lysosomes. This adaptive tuning of MAP organization in response to nutrient availability provides a mechanism for spatial reorganization of lysosomes to meet cellular metabolic demands.

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: Proposed model The schematic presents a model integrating findings from this study and prior reports , , , . Under homeostatic conditions, the microtubule network is differentially decorated with MAPs: MAP4 is enriched on tyrosinated microtubules, while MAP7D1 is enriched on detyrosinated microtubules. These MAP–PTM combinations establish specialized tracks that spatiotemporally regulate lysosome transport. On MAP7D1-decorated detyrosinated microtubules, kinesin-1 is preferentially recruited but its motility is dampened by the high density of MAP7D1 nanoclusters, promoting lysosomal immobilization in the perinuclear region to support autophagy-related functions. In contrast, MAP4-enriched tyrosinated microtubules favor fast kinesin-3 or kinesin-2–driven anterograde transport, redistributing lysosomes toward the cell periphery to enhance mTORC1 signaling. Importantly, anterograde transport is dynamically balanced by dynein-driven retrograde movement, which counteracts kinesin activity and maintains equilibrium between perinuclear and peripheral lysosome pools during homeostasis. Upon nutrient modulation, MAP densities are remodelled along microtubule tracks. Nutrient deprivation increases MAP7D1 density and reduces MAP4 abundance, thereby immobilizing kinesin-1 and biasing transport toward dynein-driven retrograde motility, which retains lysosomes in the perinuclear region. Conversely, nutrient stimulation decreases MAP7D1 density, while maintaining optimal MAP4 density, thereby enabling faster motility of kinesin and promoting anterograde transport and peripheral positioning of lysosomes. This adaptive tuning of MAP organization in response to nutrient availability provides a mechanism for spatial reorganization of lysosomes to meet cellular metabolic demands.

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

Techniques: Activity Assay

Review

human map4 (OriGene)

Structured Review

Images

1) Product Images from "Phosphatidylinositol 3-kinase Signaling is Spatially Organized at Endosomal Compartments by Microtubule-associated Protein 4"

Related Articles

Cloning:

Construct:

Variant Assay:

Synthesized:

Clone Assay:

Expressing:

Binding Assay:

Generated:

Recombinant:

Purification:

other:

Similar Products

Image Search Results