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Addgene inc mcherry cav 1
Mcherry Cav 1, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 93 stars, based on 10 article reviews

mcherry cav 1 - by Bioz Stars, 2026-05

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The association of actin filaments with cytoplasmic CAV-1. ( A ) Time-lapse imaging of U2OS cells expressing CAV-1-mEGFP and mCherry-actin revealing the motility of cytoplasmic CAV-1 vesicles along actin filaments. Two magnified regions (yellow boxes-defined Roi1 and Roi2) are chosen. White and yellow circles in the magnified regions indicate the starting and ending positions of discrete CAV-1-tagged vesicles. Scale bar, 10 µm (in cell image), 2 µm (magnified yellow box Roi 1 image), and 2 µm (magnified yellow box Roi 2 image), respectively. ( B ) The percentage of CAV-1 colocalized with actin. n = 30 cells are used for quantification. ( C ) Pearson’s coefficient of CAV-1 and actin. n = 30 cells are used for quantification. ( D ) Moving trajectories of CAV-1-positive vesicles 1, 2, and 3 in A. Red dots and green arrows indicate the starting and ending points. Scale bar, 1 μm. ( E ) The kymograph analysis of CAV-1-positive vesicles 1, 2, and 3 in A . White arrow in vesicle 3 illustrates the fast-moving trail of CAV-1 signals. The vertical scale bar represents 1 min and the horizontal scale bar represents 1 µm. ( F ) The percentage of distinct mobility types of cytoplasmic CAV-1. n = 16 cells are used for quantification. The data are presented as mean ± SEM. ( G ) The mean length of ‘dwelling’ ( n = 34 vesicles) and ‘go and dwelling’ ( n = 37 vesicles) CAV-1-positive vesicles. Data are represented as mean ± SEM. * P ≤ 0.05 ( t -test).

Journal: Journal of Molecular Cell Biology

Article Title: Actin nucleator formins regulate the tension-buffering function of caveolin-1

doi: 10.1093/jmcb/mjab070

Figure Lengend Snippet: The association of actin filaments with cytoplasmic CAV-1. ( A ) Time-lapse imaging of U2OS cells expressing CAV-1-mEGFP and mCherry-actin revealing the motility of cytoplasmic CAV-1 vesicles along actin filaments. Two magnified regions (yellow boxes-defined Roi1 and Roi2) are chosen. White and yellow circles in the magnified regions indicate the starting and ending positions of discrete CAV-1-tagged vesicles. Scale bar, 10 µm (in cell image), 2 µm (magnified yellow box Roi 1 image), and 2 µm (magnified yellow box Roi 2 image), respectively. ( B ) The percentage of CAV-1 colocalized with actin. n = 30 cells are used for quantification. ( C ) Pearson’s coefficient of CAV-1 and actin. n = 30 cells are used for quantification. ( D ) Moving trajectories of CAV-1-positive vesicles 1, 2, and 3 in A. Red dots and green arrows indicate the starting and ending points. Scale bar, 1 μm. ( E ) The kymograph analysis of CAV-1-positive vesicles 1, 2, and 3 in A . White arrow in vesicle 3 illustrates the fast-moving trail of CAV-1 signals. The vertical scale bar represents 1 min and the horizontal scale bar represents 1 µm. ( F ) The percentage of distinct mobility types of cytoplasmic CAV-1. n = 16 cells are used for quantification. The data are presented as mean ± SEM. ( G ) The mean length of ‘dwelling’ ( n = 34 vesicles) and ‘go and dwelling’ ( n = 37 vesicles) CAV-1-positive vesicles. Data are represented as mean ± SEM. * P ≤ 0.05 ( t -test).

Article Snippet: The time-lapse images of cells with transient transfection of CAV-1-mEGFP, mCherry-actin, and CAV-1-mCherry were acquired with 3I Marianas imaging system (3I intelligent Imaging Innovations), consisting of an inverted spinning disk confocal microscope Zeiss Axio Observer Z1 (Zeiss) and a Yokogawa CSU-X1 M1 confocal scanner.

Techniques: Imaging, Expressing

Inhibition of formin results in reduced density and motility of cytoplasmic CAV-1. ( A and B ) The transcriptional level of CAV-1 ( A ) and the translational level of CAV-1 and cavin-1 ( B ) are examined by real-time quantitative PCR and western blotting analysis in the wild-type U2OS (Ctrl) cells and formin-inhibited cells. GAPDH is probed for equal sample loading. ( C ) Immunofluorescence staining of endogenous F-actin and CAV-1-positive vesicles in wild-type and formin-inhibited cells. Scale bar, 10 µm. ( D ) Magnified images indicated by yellow box in C illustrate the changes of CAV-1 vesicles. Scale bar, 4 µm. The representative analysis of CAV-1-positive dots was detected by Imaris. Identified dots are marked as balls, which are randomly colored in the lower panel. The size of the color balls indicates the calculated CAV-1 vesicle size. ( E ) Quantification of the number of CAV-1-positive vesicles per µm 2 in the Ctrl ( n = 20) and formin inhibition ( n = 24) groups. The data are presented as mean ± SEM. ** P ≤ 0.01 ( t -test). ( F ) The length distribution of CAV-1-positive vesicles. The number of vesicles in each group of size is divided by the total CAV-1 number of the same cell. n = 25602 vesicles from 32 cells (Ctrl) and 12035 vesicles from 30 cells (formin inhibition). Data are represented as mean ± SEM. ( G ) The representative dot tracking analysis of CAV-1-positive vesicles in formin-inhibited cells by Imaris. White dashed line indicates the outline of the cell. Color-coded bar from blue to red indicates the tracked mean speeds ranging from 0 to 0.3 μm/sec. Scale bar, 10 μm. ( H ) Quantification of the movement rate of CAV-1-marked vesicles in Ctrl ( n = 23) and formin-inhibited ( n = 22) cells. The data are presented as mean ± SEM. *** P ≤ 0.001 ( t -test).

Journal: Journal of Molecular Cell Biology

Article Title: Actin nucleator formins regulate the tension-buffering function of caveolin-1

doi: 10.1093/jmcb/mjab070

Figure Lengend Snippet: Inhibition of formin results in reduced density and motility of cytoplasmic CAV-1. ( A and B ) The transcriptional level of CAV-1 ( A ) and the translational level of CAV-1 and cavin-1 ( B ) are examined by real-time quantitative PCR and western blotting analysis in the wild-type U2OS (Ctrl) cells and formin-inhibited cells. GAPDH is probed for equal sample loading. ( C ) Immunofluorescence staining of endogenous F-actin and CAV-1-positive vesicles in wild-type and formin-inhibited cells. Scale bar, 10 µm. ( D ) Magnified images indicated by yellow box in C illustrate the changes of CAV-1 vesicles. Scale bar, 4 µm. The representative analysis of CAV-1-positive dots was detected by Imaris. Identified dots are marked as balls, which are randomly colored in the lower panel. The size of the color balls indicates the calculated CAV-1 vesicle size. ( E ) Quantification of the number of CAV-1-positive vesicles per µm 2 in the Ctrl ( n = 20) and formin inhibition ( n = 24) groups. The data are presented as mean ± SEM. ** P ≤ 0.01 ( t -test). ( F ) The length distribution of CAV-1-positive vesicles. The number of vesicles in each group of size is divided by the total CAV-1 number of the same cell. n = 25602 vesicles from 32 cells (Ctrl) and 12035 vesicles from 30 cells (formin inhibition). Data are represented as mean ± SEM. ( G ) The representative dot tracking analysis of CAV-1-positive vesicles in formin-inhibited cells by Imaris. White dashed line indicates the outline of the cell. Color-coded bar from blue to red indicates the tracked mean speeds ranging from 0 to 0.3 μm/sec. Scale bar, 10 μm. ( H ) Quantification of the movement rate of CAV-1-marked vesicles in Ctrl ( n = 23) and formin-inhibited ( n = 22) cells. The data are presented as mean ± SEM. *** P ≤ 0.001 ( t -test).

Techniques: Inhibition, Real-time Polymerase Chain Reaction, Western Blot, Immunofluorescence, Staining

The effects of formin components FHOD1 and Dia1 on the density, size, and motility of cytoplasmic CAV-1 vesicles. ( A ) Western blotting analysis of endogenous FHOD1 and Dia1 levels in total cell lysates of wild-type (Ctrl), FHOD1 knockdown (FD1 siRNA), Dia1 knockdown (Dia1 siRNA), and FHOD1/Dia1 double knockdown (FD1 + Dia1 siRNA) U2OS cells, respectively. GAPDH is probed for equal sample loading. ( B ) Quantification of the number of CAV-1-positive vesicles per µm 2 in the Ctrl ( n = 24), FD1 siRNA ( n = 12), Dia1 siRNA ( n = 18), and FD1 + Dia1 siRNA ( n = 23) U2OS cells. The data are presented as mean ± SEM. ** P ≤ 0.01 ( t -test). ( C ) The length distribution of CAV-1-positive vesicles. The number of vesicles in each size group is divided by the total CAV-1 number of the same cell. n = 17602 vesicles from 24 cells (Ctrl), 6432 vesicles from 12 cells (FD1 siRNA), 9543 vesicles from 18 cells (Dia1 siRNA), and 10231 vesicles from 23 cells (FD1 + Dia1 siRNA). Data are represented as mean ± SEM. ( D ) Quantification of the movement rate of CAV-1-marked vesicles in the Ctrl ( n = 25), FD1 siRNA ( n = 13), Dia1 siRNA ( n = 14), and FD1 + Dia1 siRNA ( n = 24) cells. ( E ) Quantification of the ratio of ‘go and dwelling’ to ‘dwelling’ in the Ctrl ( n = 9), formin-inhibited ( n = 9), FD1 siRNA ( n = 9), and Dia1 siRNA ( n = 9) cells. The data are presented as mean ± SEM. ** P ≤ 0.01 ( t -test). ( F ) Immunofluorescence staining of endogenous F-actin and CAV-1-positive vesicles in U2OS cells expressing GFP, active GFP-FHOD1, and active GFP-Dia1, respectively. Scale bar, 10 and 5 µm in images and magnified images, respectively. ( G ) Quantification of the number of CAV-1-positive vesicles per µm 2 in each group. n = 30 cells. The data are presented as mean ± SEM. *** P ≤ 0.001 ( t -test). ( H ) The length distribution of CAV-1-positive vesicles. The number of vesicles in each size group is divided by the total CAV-1 number of the same cell. n = 16597 vesicles from 20 cells (Ctrl), 15737 vesicles from 20 cells (GFP), 20623 vesicles from 20 cells (active GFP-FHOD1), and 19405 vesicles from 20 cells (active GFP-mDia1). Data are represented as mean ± SEM.

Journal: Journal of Molecular Cell Biology

Article Title: Actin nucleator formins regulate the tension-buffering function of caveolin-1

doi: 10.1093/jmcb/mjab070

Figure Lengend Snippet: The effects of formin components FHOD1 and Dia1 on the density, size, and motility of cytoplasmic CAV-1 vesicles. ( A ) Western blotting analysis of endogenous FHOD1 and Dia1 levels in total cell lysates of wild-type (Ctrl), FHOD1 knockdown (FD1 siRNA), Dia1 knockdown (Dia1 siRNA), and FHOD1/Dia1 double knockdown (FD1 + Dia1 siRNA) U2OS cells, respectively. GAPDH is probed for equal sample loading. ( B ) Quantification of the number of CAV-1-positive vesicles per µm 2 in the Ctrl ( n = 24), FD1 siRNA ( n = 12), Dia1 siRNA ( n = 18), and FD1 + Dia1 siRNA ( n = 23) U2OS cells. The data are presented as mean ± SEM. ** P ≤ 0.01 ( t -test). ( C ) The length distribution of CAV-1-positive vesicles. The number of vesicles in each size group is divided by the total CAV-1 number of the same cell. n = 17602 vesicles from 24 cells (Ctrl), 6432 vesicles from 12 cells (FD1 siRNA), 9543 vesicles from 18 cells (Dia1 siRNA), and 10231 vesicles from 23 cells (FD1 + Dia1 siRNA). Data are represented as mean ± SEM. ( D ) Quantification of the movement rate of CAV-1-marked vesicles in the Ctrl ( n = 25), FD1 siRNA ( n = 13), Dia1 siRNA ( n = 14), and FD1 + Dia1 siRNA ( n = 24) cells. ( E ) Quantification of the ratio of ‘go and dwelling’ to ‘dwelling’ in the Ctrl ( n = 9), formin-inhibited ( n = 9), FD1 siRNA ( n = 9), and Dia1 siRNA ( n = 9) cells. The data are presented as mean ± SEM. ** P ≤ 0.01 ( t -test). ( F ) Immunofluorescence staining of endogenous F-actin and CAV-1-positive vesicles in U2OS cells expressing GFP, active GFP-FHOD1, and active GFP-Dia1, respectively. Scale bar, 10 and 5 µm in images and magnified images, respectively. ( G ) Quantification of the number of CAV-1-positive vesicles per µm 2 in each group. n = 30 cells. The data are presented as mean ± SEM. *** P ≤ 0.001 ( t -test). ( H ) The length distribution of CAV-1-positive vesicles. The number of vesicles in each size group is divided by the total CAV-1 number of the same cell. n = 16597 vesicles from 20 cells (Ctrl), 15737 vesicles from 20 cells (GFP), 20623 vesicles from 20 cells (active GFP-FHOD1), and 19405 vesicles from 20 cells (active GFP-mDia1). Data are represented as mean ± SEM.

Techniques: Western Blot, Immunofluorescence, Staining, Expressing

The linear elongated actin network by formins is critical for CAV-1 disappearance upon hypo-osmotic shock. ( A ) Time-lapse imaging of U2OS cells expressing CAV-1-mCherry cultured under routine culture condition (iso-osmosis) with formin inhibition or FHOD1 + Dia1 double knockdown, respectively. ( B ) Time-lapse imaging of U2OS cells expressing CAV-1-mCherry upon hypo-osmotic shock under Ctrl, formin inhibition, or FHOD1 + Dia1 double knockdown condition, respectively. In A and B , white dash lines indicate the outline of the cells. CAV-1-positive vesicles were detected by Imaris at different time points. Vanished CAV-1 vesicles upon 2- and 5-min iso-osmotic ( A ) or hypo-osmotic ( B ) shock are labeled by green and orange dots, respectively. Red dots illustrate the remaining CAV-1-positive vesicles after 5-min hypo-osmotic treatment. Scale bar, 10 µm. ( C ) Quantification of the percentage of CAV-1-positive vesicles left upon 5-min hypo- or iso-osmotic shock under Ctrl, formin inhibition, or FHOD1 + Dia1 double knockdown conditions, respectively. The data are presented as mean ± SEM. ** P ≤ 0.01, *** P ≤ 0.001 ( t -test). ( D ) Western blotting analysis of CAV-1, cavin-1, and actin levels in U2OS cells with formin inhibition upon hypo-osmotic shock. The blot is also probed with GAPDH antibody to verify equal sample loading. The obtained intensity value from wild-type cells was set to 1. n = 3. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way ANOVA).

Journal: Journal of Molecular Cell Biology

Article Title: Actin nucleator formins regulate the tension-buffering function of caveolin-1

doi: 10.1093/jmcb/mjab070

Figure Lengend Snippet: The linear elongated actin network by formins is critical for CAV-1 disappearance upon hypo-osmotic shock. ( A ) Time-lapse imaging of U2OS cells expressing CAV-1-mCherry cultured under routine culture condition (iso-osmosis) with formin inhibition or FHOD1 + Dia1 double knockdown, respectively. ( B ) Time-lapse imaging of U2OS cells expressing CAV-1-mCherry upon hypo-osmotic shock under Ctrl, formin inhibition, or FHOD1 + Dia1 double knockdown condition, respectively. In A and B , white dash lines indicate the outline of the cells. CAV-1-positive vesicles were detected by Imaris at different time points. Vanished CAV-1 vesicles upon 2- and 5-min iso-osmotic ( A ) or hypo-osmotic ( B ) shock are labeled by green and orange dots, respectively. Red dots illustrate the remaining CAV-1-positive vesicles after 5-min hypo-osmotic treatment. Scale bar, 10 µm. ( C ) Quantification of the percentage of CAV-1-positive vesicles left upon 5-min hypo- or iso-osmotic shock under Ctrl, formin inhibition, or FHOD1 + Dia1 double knockdown conditions, respectively. The data are presented as mean ± SEM. ** P ≤ 0.01, *** P ≤ 0.001 ( t -test). ( D ) Western blotting analysis of CAV-1, cavin-1, and actin levels in U2OS cells with formin inhibition upon hypo-osmotic shock. The blot is also probed with GAPDH antibody to verify equal sample loading. The obtained intensity value from wild-type cells was set to 1. n = 3. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way ANOVA).

Techniques: Imaging, Expressing, Cell Culture, Inhibition, Labeling, Western Blot

Inhibition of formin promotes the reduced density, increased size, and decreased motility of cytoplasmic CAV-1 vesicles when grown on soft matrix. ( A and B ) Representative live cell imaging of actin and CAV-1-positive vesicles in U2OS cells grown on glass or soft substrate 25 or 0.5 kPa, with or without formin inhibitor treatment, respectively. Scale bar, 10 µm. ( C ) Quantification of the number of CAV-1-positive vesicles per µm 2 in the cells grown on glass ( n = 29 for Ctrl, n = 22 for formin inhibition) or 25 kPa ( n = 25 for Ctrl, n = 19 for formin inhibition) and 0.5 kPa ( n = 30 for Ctrl, n = 17 for formin inhibition) substrates. The data are presented as mean ± SEM. *** P ≤ 0.001 ( t -test). ( D ) The length distribution of CAV-1-positive vesicles. The number of vesicles in each size group is divided by the total CAV-1 number of the same cell. n = 25602 vesicles from 32 Ctrl cells and 16786 vesicles from 29 formin-inhibited cells (glass), 13467 vesicles from 31 Ctrl cells and 8546 vesicles from 33 formin-inhibited cells (25 kPa), and 14567 vesicles from 32 Ctrl cells and 7658 vesicles from 34 formin-inhibited cells (0.5 kPa). Data are represented as mean ±SEM. ** P < 0.01, *** P ≤ 0.001 ( t -test). ( E ) Quantification of the movement rate of CAV-1-positive vesicles in the cells grown on glass ( n = 26 for Ctrl, n = 22 for formin inhibition) and 25 kPa ( n = 12 for Ctrl, n = 13 for formin inhibition) and 0.5 kPa ( n = 14 for Ctrl, n = 10 for formin inhibition) substrates. The data are presented as mean ± SEM. ( F ) The schematic working model. CAV-1 vesicles are associated with and can move on actin filaments in normal cells. Formin inhibition results in decreased CAV-1 vesicle numbers, enlarged vesicle areas, and slowed movement speeds. When cells are challenged with hypo-osmotic shock or softer matrix, CAV-1 vesicles become less in number, larger in area, and slower in speed in order to resist the external environment pressure. Formin inhibition expedites the changes of CAV-1 vesicles in all aspects.

Journal: Journal of Molecular Cell Biology

Article Title: Actin nucleator formins regulate the tension-buffering function of caveolin-1

doi: 10.1093/jmcb/mjab070

Figure Lengend Snippet: Inhibition of formin promotes the reduced density, increased size, and decreased motility of cytoplasmic CAV-1 vesicles when grown on soft matrix. ( A and B ) Representative live cell imaging of actin and CAV-1-positive vesicles in U2OS cells grown on glass or soft substrate 25 or 0.5 kPa, with or without formin inhibitor treatment, respectively. Scale bar, 10 µm. ( C ) Quantification of the number of CAV-1-positive vesicles per µm 2 in the cells grown on glass ( n = 29 for Ctrl, n = 22 for formin inhibition) or 25 kPa ( n = 25 for Ctrl, n = 19 for formin inhibition) and 0.5 kPa ( n = 30 for Ctrl, n = 17 for formin inhibition) substrates. The data are presented as mean ± SEM. *** P ≤ 0.001 ( t -test). ( D ) The length distribution of CAV-1-positive vesicles. The number of vesicles in each size group is divided by the total CAV-1 number of the same cell. n = 25602 vesicles from 32 Ctrl cells and 16786 vesicles from 29 formin-inhibited cells (glass), 13467 vesicles from 31 Ctrl cells and 8546 vesicles from 33 formin-inhibited cells (25 kPa), and 14567 vesicles from 32 Ctrl cells and 7658 vesicles from 34 formin-inhibited cells (0.5 kPa). Data are represented as mean ±SEM. ** P < 0.01, *** P ≤ 0.001 ( t -test). ( E ) Quantification of the movement rate of CAV-1-positive vesicles in the cells grown on glass ( n = 26 for Ctrl, n = 22 for formin inhibition) and 25 kPa ( n = 12 for Ctrl, n = 13 for formin inhibition) and 0.5 kPa ( n = 14 for Ctrl, n = 10 for formin inhibition) substrates. The data are presented as mean ± SEM. ( F ) The schematic working model. CAV-1 vesicles are associated with and can move on actin filaments in normal cells. Formin inhibition results in decreased CAV-1 vesicle numbers, enlarged vesicle areas, and slowed movement speeds. When cells are challenged with hypo-osmotic shock or softer matrix, CAV-1 vesicles become less in number, larger in area, and slower in speed in order to resist the external environment pressure. Formin inhibition expedites the changes of CAV-1 vesicles in all aspects.

Techniques: Inhibition, Live Cell Imaging

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