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monkey kidney cells (ATCC)

Name: monkey kidney cells
Brand: ATCC
Rating: 99 (9168 reviews)

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ATCC monkey kidney cells
Monkey Kidney Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 9168 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 99 stars, based on 9168 article reviews

monkey kidney cells - by Bioz Stars, 2026-02

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(a) Schematic showing the experimental plan for CgA/PA interaction monitoring in living cells by the FLIM-FRET technique. Following the overexpression of CgA- or CgAΔPABD-mKate2 in COS-7 cells, their secretion was induced during 5 min using a BaCl 2 solution. Then cells were washed and incubated 5 min with PA-ATTO647N. (b) Principle of FLIM-FRET technique to track the lifetime of the fluorescence donor (mKate2) without or with CgA/PA-ATTO647N interaction. The more CgA-mKate2 interacts with PA, the more the lifetime of mKate2 decreases due to its energy transfer to the acceptor fluorophore (ATTO647N). (c) Confocal observations of COS-7 cells overexpressing CgA-mKate2 or CgAΔPABD-mKate2 (green) after secretion stimulation and incubation with the PA-ATTO647N probe (magenta). Scale bars: 10 µm. The regions delimited by a white square on left images are enlarged to show the distribution of each fluorescent molecule. Scale bars: 2 µm. (d) Confocal observations of COS-7 cells overexpressing CgA-mKate2 or CgAΔPABD-mKate2, after cell incubation with PA-ATTO647N, showing the intensity of mKate2 fluorescence (grey), and mKate2 lifetime between 3 and 1.8 ns (rainbow color bar). The regions delimited by a white square are enlarged to show mKate2 lifetime at the plasma membrane. Scale bars: 5 µm. (e) Representative phasor plots showing the CgA-mKate2 lifetime at the level of the whole image and at the plasma membrane zone. Green line corresponds to 2.4 ns and yellow line to 2 ns, used as visual landmarks. (f) Representative phasor plot showing the CgAΔPABD-mKate2 lifetime at the level of the whole image. Green line corresponds to 2.4 ns and yellow line to 2 ns, used as visual landmarks. (g) Quantification of CgA-mKate2 or CgAΔPABD-mKate2 lifetime at the level of the plasma membrane (PM) or secretory granule (SG) without (-) or with (+) PA-ATTO647N. nd: not detected. Kruskall-Wallis test * p<0,05, each point representing the mean mKate2 lifetime in a ROI at the PM or SG level of 2 to 3 cells.

Journal: bioRxiv

Article Title: Chromogranin A regulates the dynamics of neurosecretion through its interaction with phosphatidic acid

doi: 10.64898/2026.01.29.699889

Figure Lengend Snippet: (a) Schematic showing the experimental plan for CgA/PA interaction monitoring in living cells by the FLIM-FRET technique. Following the overexpression of CgA- or CgAΔPABD-mKate2 in COS-7 cells, their secretion was induced during 5 min using a BaCl 2 solution. Then cells were washed and incubated 5 min with PA-ATTO647N. (b) Principle of FLIM-FRET technique to track the lifetime of the fluorescence donor (mKate2) without or with CgA/PA-ATTO647N interaction. The more CgA-mKate2 interacts with PA, the more the lifetime of mKate2 decreases due to its energy transfer to the acceptor fluorophore (ATTO647N). (c) Confocal observations of COS-7 cells overexpressing CgA-mKate2 or CgAΔPABD-mKate2 (green) after secretion stimulation and incubation with the PA-ATTO647N probe (magenta). Scale bars: 10 µm. The regions delimited by a white square on left images are enlarged to show the distribution of each fluorescent molecule. Scale bars: 2 µm. (d) Confocal observations of COS-7 cells overexpressing CgA-mKate2 or CgAΔPABD-mKate2, after cell incubation with PA-ATTO647N, showing the intensity of mKate2 fluorescence (grey), and mKate2 lifetime between 3 and 1.8 ns (rainbow color bar). The regions delimited by a white square are enlarged to show mKate2 lifetime at the plasma membrane. Scale bars: 5 µm. (e) Representative phasor plots showing the CgA-mKate2 lifetime at the level of the whole image and at the plasma membrane zone. Green line corresponds to 2.4 ns and yellow line to 2 ns, used as visual landmarks. (f) Representative phasor plot showing the CgAΔPABD-mKate2 lifetime at the level of the whole image. Green line corresponds to 2.4 ns and yellow line to 2 ns, used as visual landmarks. (g) Quantification of CgA-mKate2 or CgAΔPABD-mKate2 lifetime at the level of the plasma membrane (PM) or secretory granule (SG) without (-) or with (+) PA-ATTO647N. nd: not detected. Kruskall-Wallis test * p<0,05, each point representing the mean mKate2 lifetime in a ROI at the PM or SG level of 2 to 3 cells.

Article Snippet: African green monkey kidney fibroblast-derived COS-7 cells (American Type Culture Collection; CRL 1651) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Gibco, Thermo FisherScientific) supplemented with 5% fetal bovine serum (FBS, Sigma-Aldrich), 100 U/mL penicillin, 100 μg/mL streptomycin (Gibco, Thermo FisherScientific).

Techniques: Over Expression, Incubation, Fluorescence, Clinical Proteomics, Membrane

(a) TIRF-M imaging of COS-7 cells overexpressing and secreting CgA- and CgAΔPABD-pHluorin after 2 mM BaCl 2 stimulation. SG leading to exocytic events on whole cells are surrounded and indicated by yellow arrows. Scale bars: 10 µm. Below are represented typical images of CgA- or CgAΔPABD-pHluorin exocytosis events. (b) Quantification of the number of exocytosis events in COS-7 cells overexpressing CgA-pHluorin and CgAΔPABD-pHluorin after their stimulation. (n=4 independent experiments; 8 cells in CgA condition and 7 cells in CgAΔPABD condition). Data are represented as mean ± SEM. Each point is one analyzed cell and each color is an independent experiment. **p < 0.01, Mann-Whitney test. (c) Typical images of CgA-EGFP or CgAΔPABD-EGFP exocytosis events. Scale bar: 500 nm. (d) Curves represent the normalized variation of the fluorescence intensity of single exocytic events during 10 s from CgA-or CgAΔPABD-EGFP overexpressing COS-7 cells after 2 mM BaCl 2 stimulation. The mean ± SEM of 75 exocytosis of 5 different CgA-EGFP overexpressing cells in 3 independent experiments and the mean of 65 exocytic events of 7 different CgAΔPABD-EGFP overexpressing cells in 3 independent experiments. Data have been normalized between 0 (before exocytosis) and 1 (maximum intensity). (e) Plot representing the mean ± SEM of area under curves from CgA-EGFP or CgAΔPABD-EGFP secretion kinetic. Each point represents one exocytosis event, and one color represents one analyzed cell. ****p < 0.0001, Mann-Whitney test.

Journal: bioRxiv

Article Title: Chromogranin A regulates the dynamics of neurosecretion through its interaction with phosphatidic acid

doi: 10.64898/2026.01.29.699889

Figure Lengend Snippet: (a) TIRF-M imaging of COS-7 cells overexpressing and secreting CgA- and CgAΔPABD-pHluorin after 2 mM BaCl 2 stimulation. SG leading to exocytic events on whole cells are surrounded and indicated by yellow arrows. Scale bars: 10 µm. Below are represented typical images of CgA- or CgAΔPABD-pHluorin exocytosis events. (b) Quantification of the number of exocytosis events in COS-7 cells overexpressing CgA-pHluorin and CgAΔPABD-pHluorin after their stimulation. (n=4 independent experiments; 8 cells in CgA condition and 7 cells in CgAΔPABD condition). Data are represented as mean ± SEM. Each point is one analyzed cell and each color is an independent experiment. **p < 0.01, Mann-Whitney test. (c) Typical images of CgA-EGFP or CgAΔPABD-EGFP exocytosis events. Scale bar: 500 nm. (d) Curves represent the normalized variation of the fluorescence intensity of single exocytic events during 10 s from CgA-or CgAΔPABD-EGFP overexpressing COS-7 cells after 2 mM BaCl 2 stimulation. The mean ± SEM of 75 exocytosis of 5 different CgA-EGFP overexpressing cells in 3 independent experiments and the mean of 65 exocytic events of 7 different CgAΔPABD-EGFP overexpressing cells in 3 independent experiments. Data have been normalized between 0 (before exocytosis) and 1 (maximum intensity). (e) Plot representing the mean ± SEM of area under curves from CgA-EGFP or CgAΔPABD-EGFP secretion kinetic. Each point represents one exocytosis event, and one color represents one analyzed cell. ****p < 0.0001, Mann-Whitney test.

Techniques: Imaging, MANN-WHITNEY, Fluorescence

a-f . Representative images of live BODIPY (blue)-stained COS7 cells transiently transfected with GFP-ATG2A (green) and Halo-VAPs (magenta; MOSPD1/2/3, VAPA and VAPB) with insets from at least 3 independent experiments. g. In cells as in ( a-f ), Pearson’s correlation coefficient of ATG2A vs different ER proteins including Halo-Sec61β (11 cells), Halo-MOSPD1 (13 cells), Halo-MOSPD3 (13 cells), Halo-MOSPD2 (14 cells), Halo-VAPA (13 cells), or Halo-VAPB (12 cells) in three independent experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. Scale bar, 10 μm in the whole cell images and 2 μm in the insets in (a-f).

Journal: bioRxiv

Article Title: MOSPD3 mediates the ER recruitment of ATG2A to promote autophagy

doi: 10.64898/2026.01.04.697522

Figure Lengend Snippet: a-f . Representative images of live BODIPY (blue)-stained COS7 cells transiently transfected with GFP-ATG2A (green) and Halo-VAPs (magenta; MOSPD1/2/3, VAPA and VAPB) with insets from at least 3 independent experiments. g. In cells as in ( a-f ), Pearson’s correlation coefficient of ATG2A vs different ER proteins including Halo-Sec61β (11 cells), Halo-MOSPD1 (13 cells), Halo-MOSPD3 (13 cells), Halo-MOSPD2 (14 cells), Halo-VAPA (13 cells), or Halo-VAPB (12 cells) in three independent experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. Scale bar, 10 μm in the whole cell images and 2 μm in the insets in (a-f).

Article Snippet: The African green monkey kidney fibroblast-like COS7 cell line (ATCC), human cervical cancer HeLa cells (ATCC), human embryonic kidney (HEK) 293T (ATCC) and human osteosarcoma U2OS ATG2A-KI cell line were grown in DMEM (Invitrogen) supplemented with 10% fetal bovine serum (Gibco).

Techniques: Staining, Transfection

$a. Representative GFP-Trap assays demonstrating an interaction between endogenous ATG2A-GFP-KI and endogenous MOSPD3 from at least 3 independent experiments. b. Representative images of a live U2OS ATG2A-GFP KI cell transiently transfected with Halo-MOSPD3 (magenta) and mCh-LC3B (blue) with insets under EBSS stimulation. c. The percentage of endogenous ATG2A puncta that colocalized with MOSPD3 puncta (832 ATG2A-GFP puncta from 15 cells) from at least 3 independent experiments. Mean ± SD. d. Time-lapse montages of a COS7 cell transiently transfected with GFP-DFCP (green) and Halo-MOSPD3 (magenta) with white arrows denoting dynamic MOSPD3 puncta and yellow arrows denoting colocalization of GFP-DFCP1 puncta and Halo-MOSPD3 puncta from 3 independent experiments. e. The percentage of DFCP1 puncta relative to MOSPD3 puncta from 12 cells. Mean ± SD. f. The percentage of DFCP1 puncta stably associating with MOSPD3 puncta from 12 cells. GFP-DFCP1 puncta associating with Halo-MOSPD3 puncta over the imaging time window (∼15 min) were defined to be stable association. Mean ± SD. g. Membrane fractionation showing the ER association of ATG2A and ATG2B in WT and MOSPD3 KO HeLa cells in 3 independent experiments. Western blots were performed with antibodies against ATG2A, ATG2B, CANX (ER marker), Lamp1 (late endosome/PM marker), GM130 (Golgi marker) and TOM20 (mitochondrial marker). h. Membrane fractionation showing the ER association of ATG2A and ATG2B in HeLa cells treated with scrambled, VMP1, TMEM41B or VMP1+TMEM41B siRNAs in 3 independent experiments. Western blots were performed with antibodies against ATG2A, ATG2B, VMP1, TMEM41B, CANX (ER marker), Lamp1 (late endosome/PM marker), GM130 (Golgi marker) and TOM20 (mitochondrial marker). Scale bar, 10 μm in the whole cell image, 2 μm in the insets in (b, d).$

Journal: bioRxiv

Article Title: MOSPD3 mediates the ER recruitment of ATG2A to promote autophagy

doi: 10.64898/2026.01.04.697522

Figure Lengend Snippet: a. Representative GFP-Trap assays demonstrating an interaction between endogenous ATG2A-GFP-KI and endogenous MOSPD3 from at least 3 independent experiments. b. Representative images of a live U2OS ATG2A-GFP KI cell transiently transfected with Halo-MOSPD3 (magenta) and mCh-LC3B (blue) with insets under EBSS stimulation. c. The percentage of endogenous ATG2A puncta that colocalized with MOSPD3 puncta (832 ATG2A-GFP puncta from 15 cells) from at least 3 independent experiments. Mean ± SD. d. Time-lapse montages of a COS7 cell transiently transfected with GFP-DFCP (green) and Halo-MOSPD3 (magenta) with white arrows denoting dynamic MOSPD3 puncta and yellow arrows denoting colocalization of GFP-DFCP1 puncta and Halo-MOSPD3 puncta from 3 independent experiments. e. The percentage of DFCP1 puncta relative to MOSPD3 puncta from 12 cells. Mean ± SD. f. The percentage of DFCP1 puncta stably associating with MOSPD3 puncta from 12 cells. GFP-DFCP1 puncta associating with Halo-MOSPD3 puncta over the imaging time window (∼15 min) were defined to be stable association. Mean ± SD. g. Membrane fractionation showing the ER association of ATG2A and ATG2B in WT and MOSPD3 KO HeLa cells in 3 independent experiments. Western blots were performed with antibodies against ATG2A, ATG2B, CANX (ER marker), Lamp1 (late endosome/PM marker), GM130 (Golgi marker) and TOM20 (mitochondrial marker). h. Membrane fractionation showing the ER association of ATG2A and ATG2B in HeLa cells treated with scrambled, VMP1, TMEM41B or VMP1+TMEM41B siRNAs in 3 independent experiments. Western blots were performed with antibodies against ATG2A, ATG2B, VMP1, TMEM41B, CANX (ER marker), Lamp1 (late endosome/PM marker), GM130 (Golgi marker) and TOM20 (mitochondrial marker). Scale bar, 10 μm in the whole cell image, 2 μm in the insets in (b, d).

Techniques: Transfection, Stable Transfection, Imaging, Membrane, Fractionation, Western Blot, Marker

a, b . Representative images of COS7 cells expressing GFP-ATG2A (green) along with either Halo-MOSPD3 (magenta; a ) or Halo-VMP1 or TMEM41B (magenta; b ) with insets. c. In cells as in ( a, b ), Pearson’s correlation coefficient of ATG2A vs TMEM41B (10 cells), VMP1 (10 cells) or MOSPD3 (10 cells) in three independent experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. d. Representative images of COS7 cells expressing GFP-ATG2A (green) along with either Halo-VMP1 (magenta; top panel) or TMEM41B (magenta; bottom panel) with insets upon EBSS starvation from three independent experiments. e. Representative images of COS7 cells expressing GFP-ATG2A-NT (green) with Halo-MOSPD3, Halo-VMP1, or TMEM41B-Halo (magenta) with insets. f. In cells as in ( e ), Pearson’s correlation coefficient of ATG2A-NT proteins vs VMP1 (10 cells), TMEM41B (10 cells), or MOSPD3 (10 cells) from three independent experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. Scale bar, 10 μm in the whole cell images and 2 μm in the insets in (a, b, d & e).

Journal: bioRxiv

Article Title: MOSPD3 mediates the ER recruitment of ATG2A to promote autophagy

doi: 10.64898/2026.01.04.697522

Figure Lengend Snippet: a, b . Representative images of COS7 cells expressing GFP-ATG2A (green) along with either Halo-MOSPD3 (magenta; a ) or Halo-VMP1 or TMEM41B (magenta; b ) with insets. c. In cells as in ( a, b ), Pearson’s correlation coefficient of ATG2A vs TMEM41B (10 cells), VMP1 (10 cells) or MOSPD3 (10 cells) in three independent experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. d. Representative images of COS7 cells expressing GFP-ATG2A (green) along with either Halo-VMP1 (magenta; top panel) or TMEM41B (magenta; bottom panel) with insets upon EBSS starvation from three independent experiments. e. Representative images of COS7 cells expressing GFP-ATG2A-NT (green) with Halo-MOSPD3, Halo-VMP1, or TMEM41B-Halo (magenta) with insets. f. In cells as in ( e ), Pearson’s correlation coefficient of ATG2A-NT proteins vs VMP1 (10 cells), TMEM41B (10 cells), or MOSPD3 (10 cells) from three independent experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. Scale bar, 10 μm in the whole cell images and 2 μm in the insets in (a, b, d & e).

Techniques: Expressing

$a. Representative images of a COS7 cell expressing GFP-ATG2A-ΔCT (green) and Halo-MOSPD3 (magenta) with two insets from at least 3 independent experiments. Yellow asterisks mark cells expressing both GFP-ATG2A-ΔCT and Halo-MOSPD3. b. Representative images of a COS7cell expressing GFP-ATG2A-NT (green) and Halo-MOSPD3 (magenta) with two insets from at least 3 independent experiments. Yellow asterisks mark cells transfected with both GFP-ATG2A-ΔCT and Halo-MOSPD3. c. In vitro pulldown assays using purified GFP-MOSPD3 and purified 14×His-ATG2A-NT or 14×His-ATG2B-NT from 3 independent experiments. d. AlphaFold Multimer prediction of the binding sites between ATG2A and MOSPD3. e. Conservation of the FFNT motif in ATG2A across species. f. The conserved FFNT motif within ATG2A is compared with homologous regions from the indicated human proteins (ANKLE2, CERT, VPS13D, and STARD3). Conserved residues are highlighted. g. Representative images of COS7 cells expressing GFP-ATG2A-NT (green; WT, T362A, T362D and the ΔFFN mutant) and Halo-MOSPD3 (magenta) with insets from at least 3 independent experiments. h. Pearson’s correlation coefficient of MOSPD3 vs. WT ATG2A-NT (10 cells), ATG2A-NT-ΔFFNT (11 cells), ATG2A-T362A (12 cells), or ATG2A-T362D (12 cells) in three independent experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. i. Membrane fractionation showing the ER association of WT GFP-ATG2A-NT, GFP-ATG2A-NT-T362A and GFP-ATG2A-NT-T362D from 3 independent experiments. Western blots were performed with antibodies against GFP, CANX (ER marker), Lamp1 (late endosome/PM marker), GM130 (Golgi marker) and TOM20 (mitochondrial marker). Scale bar, 10 μm in the whole cell images and 2 μm in the insets in (a, b, & g).$

Journal: bioRxiv

Article Title: MOSPD3 mediates the ER recruitment of ATG2A to promote autophagy

doi: 10.64898/2026.01.04.697522

Figure Lengend Snippet: a. Representative images of a COS7 cell expressing GFP-ATG2A-ΔCT (green) and Halo-MOSPD3 (magenta) with two insets from at least 3 independent experiments. Yellow asterisks mark cells expressing both GFP-ATG2A-ΔCT and Halo-MOSPD3. b. Representative images of a COS7cell expressing GFP-ATG2A-NT (green) and Halo-MOSPD3 (magenta) with two insets from at least 3 independent experiments. Yellow asterisks mark cells transfected with both GFP-ATG2A-ΔCT and Halo-MOSPD3. c. In vitro pulldown assays using purified GFP-MOSPD3 and purified 14×His-ATG2A-NT or 14×His-ATG2B-NT from 3 independent experiments. d. AlphaFold Multimer prediction of the binding sites between ATG2A and MOSPD3. e. Conservation of the FFNT motif in ATG2A across species. f. The conserved FFNT motif within ATG2A is compared with homologous regions from the indicated human proteins (ANKLE2, CERT, VPS13D, and STARD3). Conserved residues are highlighted. g. Representative images of COS7 cells expressing GFP-ATG2A-NT (green; WT, T362A, T362D and the ΔFFN mutant) and Halo-MOSPD3 (magenta) with insets from at least 3 independent experiments. h. Pearson’s correlation coefficient of MOSPD3 vs. WT ATG2A-NT (10 cells), ATG2A-NT-ΔFFNT (11 cells), ATG2A-T362A (12 cells), or ATG2A-T362D (12 cells) in three independent experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. i. Membrane fractionation showing the ER association of WT GFP-ATG2A-NT, GFP-ATG2A-NT-T362A and GFP-ATG2A-NT-T362D from 3 independent experiments. Western blots were performed with antibodies against GFP, CANX (ER marker), Lamp1 (late endosome/PM marker), GM130 (Golgi marker) and TOM20 (mitochondrial marker). Scale bar, 10 μm in the whole cell images and 2 μm in the insets in (a, b, & g).

Techniques: Expressing, Transfection, In Vitro, Purification, Binding Assay, Mutagenesis, Membrane, Fractionation, Western Blot, Marker

a. Top: domain organization of MOSPD3. Bottom: representative images of a COS7 cell expressing GFP-ATG2A-NT (green) and Halo-MOSPD3 (magenta) with insets from at least 3 independent experiments. b. Representative images of a COS7 cell expressing GFP-ATG2A (green) and Halo-MOSPD3-ΔTM (magenta) with insets from at least 3 independent experiments. c. Representative images of a COS7 cell expressing GFP-ATG2A (green) and Halo-MOSPD3-MSP (magenta) with insets from at least 3 independent experiments. d. Representative images of a COS7 cell expressing either GFP-ATG2A-NT (green; top panel) or GFP-ATG2A (green; bottom panel) along with Halo-MOSPD3-ΔMSP (magenta) with insets from at least 3 independent experiments. e. Representative images of a COS7 cell expressing GFP-ATG2A-NT (green) and Halo-MOSPD3-ΔNT (magenta) with insets from at least 3 independent experiments. f. Representative images of a COS7 cell expressing GFP-ATG2A-NT (green) and Halo-MOSPD3-ΔPro (magenta) with insets from at least 3 independent experiments. g. In cells as in ( a-f ), Pearson’s correlation coefficient of ATG2A vs different MOSPD3 mutants including WT Halo-MOSPD3, Halo-MOSPD3-ΔNT, Halo-MOSPD3-Δ Pro, Halo-MOSPD3-ΔMSP, Halo-MOSPD3-ΔTM, Halo-MOSPD3-MSP. 10 cells for each group were analysed from three independent experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. Scale bar, 10 μm in the whole cell images and 2 μm in the insets in (a-f).

Journal: bioRxiv

Article Title: MOSPD3 mediates the ER recruitment of ATG2A to promote autophagy

doi: 10.64898/2026.01.04.697522

Figure Lengend Snippet: a. Top: domain organization of MOSPD3. Bottom: representative images of a COS7 cell expressing GFP-ATG2A-NT (green) and Halo-MOSPD3 (magenta) with insets from at least 3 independent experiments. b. Representative images of a COS7 cell expressing GFP-ATG2A (green) and Halo-MOSPD3-ΔTM (magenta) with insets from at least 3 independent experiments. c. Representative images of a COS7 cell expressing GFP-ATG2A (green) and Halo-MOSPD3-MSP (magenta) with insets from at least 3 independent experiments. d. Representative images of a COS7 cell expressing either GFP-ATG2A-NT (green; top panel) or GFP-ATG2A (green; bottom panel) along with Halo-MOSPD3-ΔMSP (magenta) with insets from at least 3 independent experiments. e. Representative images of a COS7 cell expressing GFP-ATG2A-NT (green) and Halo-MOSPD3-ΔNT (magenta) with insets from at least 3 independent experiments. f. Representative images of a COS7 cell expressing GFP-ATG2A-NT (green) and Halo-MOSPD3-ΔPro (magenta) with insets from at least 3 independent experiments. g. In cells as in ( a-f ), Pearson’s correlation coefficient of ATG2A vs different MOSPD3 mutants including WT Halo-MOSPD3, Halo-MOSPD3-ΔNT, Halo-MOSPD3-Δ Pro, Halo-MOSPD3-ΔMSP, Halo-MOSPD3-ΔTM, Halo-MOSPD3-MSP. 10 cells for each group were analysed from three independent experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. Scale bar, 10 μm in the whole cell images and 2 μm in the insets in (a-f).

Techniques: Expressing

$a. Representative images of COS7 cells transiently transfected with GFP-ATG2B (green) along with MOSPD1, MOSPD3, VAPA, VAPB or MOSPD2 (magenta) with an inset on the bottom from at least 3 independent experiments. b. In cells as in ( a ), Pearson’s correlation coefficient of ATG2B vs different VAP-related proteins including Halo-MOSPD1 (13 cells), Halo-MOSPD3 (14 cells), Halo-MOSPD2 (12 cells), Halo-VAPA (10 cells), or Halo-VAPB (11 cells) in three independent experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. c. AlphaFold Multimer prediction of the binding sites between ATG2B and MOSPD1 (left panel) or MOSPD3 (right panel). d. Conservation of the FFNT motif in ATG2B across species. e. The conserved FFNT motif within ATG2B is compared with homologous regions from the indicated human proteins (ATG2A, ANKLE2, ENTR1, CERT, VPS13D, and STARD3). Conserved residues are highlighted. f. GFP-Trap assays demonstrate interactions between ATG2B and MOSPD1/3 in HEK293 cells from three independent assays. g. Representative images of COS7 cells expressing GFP-ATG2B-ΔFFNT (green) along with either Halo-MOSPD1 (magenta; top panel) or Halo-MOSPD3 (magenta; bottom panel) with insets from at least 3 independent experiments. h. In cells as in ( g ), Pearson’s correlation coefficient of ATG2B-ΔFFNT vs Halo-MOSPD1 (12 cells), or Halo-MOSPD3 (13 cells) in three independent experiments. Unpaired student t test. Mean ± SD. i. Membrane fractionation showing the ER association of ATG2A and ATG2B in MOSPD1/3 double KO HeLa cells from 3 independent experiments. Western blots were performed with antibodies against ATG2A, ATG2B, MOSPD3, CANX (ER marker), Lamp1 (late endosome/PM marker), GM130 (Golgi marker) and TOM20 (mitochondrial marker). Scale bar, 10 μm in the whole cell images and 2 μm in the insets in (a & g).$

Journal: bioRxiv

Article Title: MOSPD3 mediates the ER recruitment of ATG2A to promote autophagy

doi: 10.64898/2026.01.04.697522

Figure Lengend Snippet: a. Representative images of COS7 cells transiently transfected with GFP-ATG2B (green) along with MOSPD1, MOSPD3, VAPA, VAPB or MOSPD2 (magenta) with an inset on the bottom from at least 3 independent experiments. b. In cells as in ( a ), Pearson’s correlation coefficient of ATG2B vs different VAP-related proteins including Halo-MOSPD1 (13 cells), Halo-MOSPD3 (14 cells), Halo-MOSPD2 (12 cells), Halo-VAPA (10 cells), or Halo-VAPB (11 cells) in three independent experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. c. AlphaFold Multimer prediction of the binding sites between ATG2B and MOSPD1 (left panel) or MOSPD3 (right panel). d. Conservation of the FFNT motif in ATG2B across species. e. The conserved FFNT motif within ATG2B is compared with homologous regions from the indicated human proteins (ATG2A, ANKLE2, ENTR1, CERT, VPS13D, and STARD3). Conserved residues are highlighted. f. GFP-Trap assays demonstrate interactions between ATG2B and MOSPD1/3 in HEK293 cells from three independent assays. g. Representative images of COS7 cells expressing GFP-ATG2B-ΔFFNT (green) along with either Halo-MOSPD1 (magenta; top panel) or Halo-MOSPD3 (magenta; bottom panel) with insets from at least 3 independent experiments. h. In cells as in ( g ), Pearson’s correlation coefficient of ATG2B-ΔFFNT vs Halo-MOSPD1 (12 cells), or Halo-MOSPD3 (13 cells) in three independent experiments. Unpaired student t test. Mean ± SD. i. Membrane fractionation showing the ER association of ATG2A and ATG2B in MOSPD1/3 double KO HeLa cells from 3 independent experiments. Western blots were performed with antibodies against ATG2A, ATG2B, MOSPD3, CANX (ER marker), Lamp1 (late endosome/PM marker), GM130 (Golgi marker) and TOM20 (mitochondrial marker). Scale bar, 10 μm in the whole cell images and 2 μm in the insets in (a & g).

Techniques: Transfection, Binding Assay, Expressing, Membrane, Fractionation, Western Blot, Marker

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