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pacgfp n1  (TaKaRa)


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    TaKaRa pacgfp n1
    Pacgfp N1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 95/100, based on 217 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pacgfp+n1/pAcGFP1-N1+Vector/pmc13134964-290-27-28
    Average 95 stars, based on 217 article reviews
    pacgfp n1 - by Bioz Stars, 2026-09
    95/100 stars

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    Related Articles

    Construct:

    Article Title: A viral APOBEC3 antagonist distinguishes HHV-6A from HHV-6B
    Article Snippet: Plasmids encoding AcGFP-A3A to -A3H were constructed by inserting CBMC cDNA or pCMV-hA3B-BE3 (Addgene plasmid #113411) into pAcGFP-N1. .. Plasmids pAcGFP-A3CH-L4, A3HC-L4, A3CH-L1, and A3CH-B1, carrying the fusion proteins of the A3C-A3H chimeras with AcGFP, were constructed by amplifying from pAcGFP-A3C or pAcGFP-A3H and cloning into pAcGFP-N1 (Clontech, 632469). ..

    Article Title: Recruitment of the SNX17-Retriever recycling pathway regulates synaptic function and plasticity
    Article Snippet: To knock-down ITGB1, a pGIPZ vector with shRNA target sequence TTCTTTATAGTTTGAGAGC (V2LMM_39157, Horizon Discovery) was utilized. .. MISSION pLKO.1 scrambled non-target shRNA SHC002 (Millipore Sigma) was used as a control for SNX17 knockdown constructs, and pGIPZ scrambled non-target (RHS4346, Horizon Discovery) was the control for the shRNAs to VPS26C and ITGB1. pmCherry-C1 (cat. No. 632524) and pAcGFP-N1 (cat. No. 632469) were purchased from Clontech. dsRed-EEA1-FYVE was a generous gift from Dr. Daniel D. Billadeau (Mayo Clinic) and has been previously described in ( ). pRK5-HA-CaMKII-T286D was a generous gift from Dr. Gentry Patrick (University of California San Diego). pCDNA4:TO was purchased from Invitrogen, and was used to generate the pCDNA4:TO-GFP_NoStop vector (without a stop codon after GFP). .. To build this vector, GFP was amplified from pAcGFP-N1 with oligos 5 ′ - AAAGGTACCATGGTGAGCAAGGGCGAGGAG-3 ′ and 5 ′ - AAGGGCCCTTACTTGTACAGCTCGTCC-3 ′ .

    Article Title: Identification and Characterization of Alternative Promoters, Transcripts and Protein Isoforms of Zebrafish R2 Gene
    Article Snippet: .. To construct vectors for ectopic expression of R1 and R2, the coding sequence of R1 gene was inserted into the vector pCGN-HAM and pDsRed1-N1 from Clontech, and coding sequences for three putative R2 isoforms were inserted into the pCMV-Tag2c and pAcGFP-N1 from Clontech, respectively. ..

    Article Title: Recruitment of the SNX17-Retriever recycling pathway regulates synaptic function and plasticity.
    Article Snippet: .. MISSION pLKO.1 scrambled nontarget shRNA SHC002 (Millipore Sigma) was used as a control for SNX17 knockdown constructs, and pGIPZ scrambled non-target (RHS4346; Horizon Discovery) was the control for the shRNAs to VPS26C and ITGB1. pmCherry-C1 (cat. no. 632524) and pAcGFP-N1 (cat. no. 632469) were purchased from Clontech. dsRed-EEA1-FYVE was a generous gift from Dr. Daniel D. Billadeau (Mayo Clinic, Rochester, MN, USA) and has been previously described in Singla et al. (2019). pRK5-HA-CaMKII-T286D was a generous gift from Dr. Gentry Patrick (University of California at San Diego, San Diego, CA, USA). pCDNA4:TO was purchased from Invitrogen and was used to generate the pCDNA4:TO-GFP_NoStop vector (without a stop codon after GFP). .. To build this vector, GFP was amplified from pAcGFP-N1 with oligos 59-AAAGGTACCATGGTGAGCAAGGGC GAGGAG-39 and 59-AAGGGCCCTTACTTGTACAGCTCGTCC-39.

    Cloning:

    Article Title: A viral APOBEC3 antagonist distinguishes HHV-6A from HHV-6B
    Article Snippet: Plasmids encoding AcGFP-A3A to -A3H were constructed by inserting CBMC cDNA or pCMV-hA3B-BE3 (Addgene plasmid #113411) into pAcGFP-N1. .. Plasmids pAcGFP-A3CH-L4, A3HC-L4, A3CH-L1, and A3CH-B1, carrying the fusion proteins of the A3C-A3H chimeras with AcGFP, were constructed by amplifying from pAcGFP-A3C or pAcGFP-A3H and cloning into pAcGFP-N1 (Clontech, 632469). ..

    shRNA:

    Article Title: Recruitment of the SNX17-Retriever recycling pathway regulates synaptic function and plasticity
    Article Snippet: To knock-down ITGB1, a pGIPZ vector with shRNA target sequence TTCTTTATAGTTTGAGAGC (V2LMM_39157, Horizon Discovery) was utilized. .. MISSION pLKO.1 scrambled non-target shRNA SHC002 (Millipore Sigma) was used as a control for SNX17 knockdown constructs, and pGIPZ scrambled non-target (RHS4346, Horizon Discovery) was the control for the shRNAs to VPS26C and ITGB1. pmCherry-C1 (cat. No. 632524) and pAcGFP-N1 (cat. No. 632469) were purchased from Clontech. dsRed-EEA1-FYVE was a generous gift from Dr. Daniel D. Billadeau (Mayo Clinic) and has been previously described in ( ). pRK5-HA-CaMKII-T286D was a generous gift from Dr. Gentry Patrick (University of California San Diego). pCDNA4:TO was purchased from Invitrogen, and was used to generate the pCDNA4:TO-GFP_NoStop vector (without a stop codon after GFP). .. To build this vector, GFP was amplified from pAcGFP-N1 with oligos 5 ′ - AAAGGTACCATGGTGAGCAAGGGCGAGGAG-3 ′ and 5 ′ - AAGGGCCCTTACTTGTACAGCTCGTCC-3 ′ .

    Article Title: Recruitment of the SNX17-Retriever recycling pathway regulates synaptic function and plasticity.
    Article Snippet: .. MISSION pLKO.1 scrambled nontarget shRNA SHC002 (Millipore Sigma) was used as a control for SNX17 knockdown constructs, and pGIPZ scrambled non-target (RHS4346; Horizon Discovery) was the control for the shRNAs to VPS26C and ITGB1. pmCherry-C1 (cat. no. 632524) and pAcGFP-N1 (cat. no. 632469) were purchased from Clontech. dsRed-EEA1-FYVE was a generous gift from Dr. Daniel D. Billadeau (Mayo Clinic, Rochester, MN, USA) and has been previously described in Singla et al. (2019). pRK5-HA-CaMKII-T286D was a generous gift from Dr. Gentry Patrick (University of California at San Diego, San Diego, CA, USA). pCDNA4:TO was purchased from Invitrogen and was used to generate the pCDNA4:TO-GFP_NoStop vector (without a stop codon after GFP). .. To build this vector, GFP was amplified from pAcGFP-N1 with oligos 59-AAAGGTACCATGGTGAGCAAGGGC GAGGAG-39 and 59-AAGGGCCCTTACTTGTACAGCTCGTCC-39.

    Control:

    Article Title: Recruitment of the SNX17-Retriever recycling pathway regulates synaptic function and plasticity
    Article Snippet: To knock-down ITGB1, a pGIPZ vector with shRNA target sequence TTCTTTATAGTTTGAGAGC (V2LMM_39157, Horizon Discovery) was utilized. .. MISSION pLKO.1 scrambled non-target shRNA SHC002 (Millipore Sigma) was used as a control for SNX17 knockdown constructs, and pGIPZ scrambled non-target (RHS4346, Horizon Discovery) was the control for the shRNAs to VPS26C and ITGB1. pmCherry-C1 (cat. No. 632524) and pAcGFP-N1 (cat. No. 632469) were purchased from Clontech. dsRed-EEA1-FYVE was a generous gift from Dr. Daniel D. Billadeau (Mayo Clinic) and has been previously described in ( ). pRK5-HA-CaMKII-T286D was a generous gift from Dr. Gentry Patrick (University of California San Diego). pCDNA4:TO was purchased from Invitrogen, and was used to generate the pCDNA4:TO-GFP_NoStop vector (without a stop codon after GFP). .. To build this vector, GFP was amplified from pAcGFP-N1 with oligos 5 ′ - AAAGGTACCATGGTGAGCAAGGGCGAGGAG-3 ′ and 5 ′ - AAGGGCCCTTACTTGTACAGCTCGTCC-3 ′ .

    Article Title: Recruitment of the SNX17-Retriever recycling pathway regulates synaptic function and plasticity.
    Article Snippet: .. MISSION pLKO.1 scrambled nontarget shRNA SHC002 (Millipore Sigma) was used as a control for SNX17 knockdown constructs, and pGIPZ scrambled non-target (RHS4346; Horizon Discovery) was the control for the shRNAs to VPS26C and ITGB1. pmCherry-C1 (cat. no. 632524) and pAcGFP-N1 (cat. no. 632469) were purchased from Clontech. dsRed-EEA1-FYVE was a generous gift from Dr. Daniel D. Billadeau (Mayo Clinic, Rochester, MN, USA) and has been previously described in Singla et al. (2019). pRK5-HA-CaMKII-T286D was a generous gift from Dr. Gentry Patrick (University of California at San Diego, San Diego, CA, USA). pCDNA4:TO was purchased from Invitrogen and was used to generate the pCDNA4:TO-GFP_NoStop vector (without a stop codon after GFP). .. To build this vector, GFP was amplified from pAcGFP-N1 with oligos 59-AAAGGTACCATGGTGAGCAAGGGC GAGGAG-39 and 59-AAGGGCCCTTACTTGTACAGCTCGTCC-39.

    Knockdown:

    Article Title: Recruitment of the SNX17-Retriever recycling pathway regulates synaptic function and plasticity
    Article Snippet: To knock-down ITGB1, a pGIPZ vector with shRNA target sequence TTCTTTATAGTTTGAGAGC (V2LMM_39157, Horizon Discovery) was utilized. .. MISSION pLKO.1 scrambled non-target shRNA SHC002 (Millipore Sigma) was used as a control for SNX17 knockdown constructs, and pGIPZ scrambled non-target (RHS4346, Horizon Discovery) was the control for the shRNAs to VPS26C and ITGB1. pmCherry-C1 (cat. No. 632524) and pAcGFP-N1 (cat. No. 632469) were purchased from Clontech. dsRed-EEA1-FYVE was a generous gift from Dr. Daniel D. Billadeau (Mayo Clinic) and has been previously described in ( ). pRK5-HA-CaMKII-T286D was a generous gift from Dr. Gentry Patrick (University of California San Diego). pCDNA4:TO was purchased from Invitrogen, and was used to generate the pCDNA4:TO-GFP_NoStop vector (without a stop codon after GFP). .. To build this vector, GFP was amplified from pAcGFP-N1 with oligos 5 ′ - AAAGGTACCATGGTGAGCAAGGGCGAGGAG-3 ′ and 5 ′ - AAGGGCCCTTACTTGTACAGCTCGTCC-3 ′ .

    Article Title: Recruitment of the SNX17-Retriever recycling pathway regulates synaptic function and plasticity.
    Article Snippet: .. MISSION pLKO.1 scrambled nontarget shRNA SHC002 (Millipore Sigma) was used as a control for SNX17 knockdown constructs, and pGIPZ scrambled non-target (RHS4346; Horizon Discovery) was the control for the shRNAs to VPS26C and ITGB1. pmCherry-C1 (cat. no. 632524) and pAcGFP-N1 (cat. no. 632469) were purchased from Clontech. dsRed-EEA1-FYVE was a generous gift from Dr. Daniel D. Billadeau (Mayo Clinic, Rochester, MN, USA) and has been previously described in Singla et al. (2019). pRK5-HA-CaMKII-T286D was a generous gift from Dr. Gentry Patrick (University of California at San Diego, San Diego, CA, USA). pCDNA4:TO was purchased from Invitrogen and was used to generate the pCDNA4:TO-GFP_NoStop vector (without a stop codon after GFP). .. To build this vector, GFP was amplified from pAcGFP-N1 with oligos 59-AAAGGTACCATGGTGAGCAAGGGC GAGGAG-39 and 59-AAGGGCCCTTACTTGTACAGCTCGTCC-39.

    Plasmid Preparation:

    Article Title: Recruitment of the SNX17-Retriever recycling pathway regulates synaptic function and plasticity
    Article Snippet: To knock-down ITGB1, a pGIPZ vector with shRNA target sequence TTCTTTATAGTTTGAGAGC (V2LMM_39157, Horizon Discovery) was utilized. .. MISSION pLKO.1 scrambled non-target shRNA SHC002 (Millipore Sigma) was used as a control for SNX17 knockdown constructs, and pGIPZ scrambled non-target (RHS4346, Horizon Discovery) was the control for the shRNAs to VPS26C and ITGB1. pmCherry-C1 (cat. No. 632524) and pAcGFP-N1 (cat. No. 632469) were purchased from Clontech. dsRed-EEA1-FYVE was a generous gift from Dr. Daniel D. Billadeau (Mayo Clinic) and has been previously described in ( ). pRK5-HA-CaMKII-T286D was a generous gift from Dr. Gentry Patrick (University of California San Diego). pCDNA4:TO was purchased from Invitrogen, and was used to generate the pCDNA4:TO-GFP_NoStop vector (without a stop codon after GFP). .. To build this vector, GFP was amplified from pAcGFP-N1 with oligos 5 ′ - AAAGGTACCATGGTGAGCAAGGGCGAGGAG-3 ′ and 5 ′ - AAGGGCCCTTACTTGTACAGCTCGTCC-3 ′ .

    Article Title: Recruitment of the SNX17-Retriever recycling pathway regulates synaptic function and plasticity.
    Article Snippet: .. MISSION pLKO.1 scrambled nontarget shRNA SHC002 (Millipore Sigma) was used as a control for SNX17 knockdown constructs, and pGIPZ scrambled non-target (RHS4346; Horizon Discovery) was the control for the shRNAs to VPS26C and ITGB1. pmCherry-C1 (cat. no. 632524) and pAcGFP-N1 (cat. no. 632469) were purchased from Clontech. dsRed-EEA1-FYVE was a generous gift from Dr. Daniel D. Billadeau (Mayo Clinic, Rochester, MN, USA) and has been previously described in Singla et al. (2019). pRK5-HA-CaMKII-T286D was a generous gift from Dr. Gentry Patrick (University of California at San Diego, San Diego, CA, USA). pCDNA4:TO was purchased from Invitrogen and was used to generate the pCDNA4:TO-GFP_NoStop vector (without a stop codon after GFP). .. To build this vector, GFP was amplified from pAcGFP-N1 with oligos 59-AAAGGTACCATGGTGAGCAAGGGC GAGGAG-39 and 59-AAGGGCCCTTACTTGTACAGCTCGTCC-39.

    Transfection:

    Article Title: Mitochondrial Lon protease is a gatekeeper for proteins newly imported into the matrix
    Article Snippet: .. A day after the second siRNA transfection, cotransfection of pDsRed2-Mito Vector (Clontech) with pAcGFP-N1 (Clontech) in a 1:1 ratio was performed using Lipofectamine LTX with the Plus Reagent according to the manufacturer’s instructions. ..

    Article Title: TFEB overexpression alleviates autophagy-lysosomal deficits caused by progranulin insufficiency
    Article Snippet: Control and GRN KO cells were plated at uniform density onto 24-well culture plates (Corning Life Sciences) and analyzed for autophagic flux four days later. .. For analysis of the effects of progranulin or TFEB on autophagy, cells were transfected with the GFP or progranulin plasmids described above, or p-EGFP-N1-TFEB (Addgene #38119) or pAcGFP-N1 (Takara # 632469) and analyzed four days after transfection. .. To analyze autophagic flux, cells were incubated in culture media with 50 μM chloroquine (MP Biomedicals) or vehicle (1:1000 sterile water diluted in media) for one hour.

    Cotransfection:

    Article Title: Mitochondrial Lon protease is a gatekeeper for proteins newly imported into the matrix
    Article Snippet: .. A day after the second siRNA transfection, cotransfection of pDsRed2-Mito Vector (Clontech) with pAcGFP-N1 (Clontech) in a 1:1 ratio was performed using Lipofectamine LTX with the Plus Reagent according to the manufacturer’s instructions. ..

    Isolation:

    Article Title: Cryptotanshinone is a candidate therapeutic agent for interstitial lung disease associated with a BRICHOS-domain mutation of SFTPC
    Article Snippet: .. AcGFP was isolated from pAcGFP-N1 (Takara) using PCR with the primers (KP051 and KP052). .. HEK293 cells were plated on 96- or 24-well PureCoat amine-coated plates (Falcon; BD Biosciences, Franklin Lakes, NJ, USA).

    Polymerase Chain Reaction:

    Article Title: Cryptotanshinone is a candidate therapeutic agent for interstitial lung disease associated with a BRICHOS-domain mutation of SFTPC
    Article Snippet: .. AcGFP was isolated from pAcGFP-N1 (Takara) using PCR with the primers (KP051 and KP052). .. HEK293 cells were plated on 96- or 24-well PureCoat amine-coated plates (Falcon; BD Biosciences, Franklin Lakes, NJ, USA).

    Expressing:

    Article Title: Identification and Characterization of Alternative Promoters, Transcripts and Protein Isoforms of Zebrafish R2 Gene
    Article Snippet: .. To construct vectors for ectopic expression of R1 and R2, the coding sequence of R1 gene was inserted into the vector pCGN-HAM and pDsRed1-N1 from Clontech, and coding sequences for three putative R2 isoforms were inserted into the pCMV-Tag2c and pAcGFP-N1 from Clontech, respectively. ..

    Sequencing:

    Article Title: Identification and Characterization of Alternative Promoters, Transcripts and Protein Isoforms of Zebrafish R2 Gene
    Article Snippet: .. To construct vectors for ectopic expression of R1 and R2, the coding sequence of R1 gene was inserted into the vector pCGN-HAM and pDsRed1-N1 from Clontech, and coding sequences for three putative R2 isoforms were inserted into the pCMV-Tag2c and pAcGFP-N1 from Clontech, respectively. ..

    Variant Assay:

    Article Title: Cooperation between Intrinsically Disordered Regions regulates CBP condensate behaviour
    Article Snippet: .. To generate human CBP wt (transcript variant 1, NCBI Reference: NM_004380.3) tagged with a C-terminal GFP for overexpression experiments (CBP wt -GFP), Homo sapiens CBP transcript variant 2 (NCBI Reference NM_001079846.1, Sino Biological HG17295-UT) was cloned in-frame into pAcGFP-N1 (Clontech) using NEBuilder HiFi DNA Assembly reaction protocol (NEB). ..

    Over Expression:

    Article Title: Cooperation between Intrinsically Disordered Regions regulates CBP condensate behaviour
    Article Snippet: .. To generate human CBP wt (transcript variant 1, NCBI Reference: NM_004380.3) tagged with a C-terminal GFP for overexpression experiments (CBP wt -GFP), Homo sapiens CBP transcript variant 2 (NCBI Reference NM_001079846.1, Sino Biological HG17295-UT) was cloned in-frame into pAcGFP-N1 (Clontech) using NEBuilder HiFi DNA Assembly reaction protocol (NEB). ..

    Clone Assay:

    Article Title: Cooperation between Intrinsically Disordered Regions regulates CBP condensate behaviour
    Article Snippet: .. To generate human CBP wt (transcript variant 1, NCBI Reference: NM_004380.3) tagged with a C-terminal GFP for overexpression experiments (CBP wt -GFP), Homo sapiens CBP transcript variant 2 (NCBI Reference NM_001079846.1, Sino Biological HG17295-UT) was cloned in-frame into pAcGFP-N1 (Clontech) using NEBuilder HiFi DNA Assembly reaction protocol (NEB). ..



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    a , Top-down (top) and oblique angle (middle, bottom) views of still frames from time-lapse 3D SoRa SDCM imaging of LAP2β-AcGFP1 (green) and SPY555_FastAct_X (magenta) in the U2OS cell region confined in a Matrigel/laminin-filled 8 μm-wide pore. Images were rendered and reconstructed in 3D for each time point. Angled side views are more zoomed in on the nuclear envelope, which appears to surround the pore. Nuclear envelope creases and folds (arrows) are seen in the 60 and 120 minute time frames. Note that the nuclear envelope preserves its continuity in the 120 minute timeframe, but the front side becomes dim due to higher signal intensity in the creased regions of the 3D-rendered image. Scale bars, 2 μm. b, Time-lapse frames of a single optical slice from SoRa SDCM imaging of a U2OS cell region confined in a Matrigel/laminin-filled 8 μm-wide pore. U2OS cells were transfected with GFP-SEPT9_i1 (green) and labeled live with SPY650_FastAct_X (magents). White arrows point to actin that colocalizes with GFP-SEPT9_i1, and enlarges overtime. Blue arrows point to actoseptin that appears in frame de novo and remains spatially stable overtime. Scale bars, 2 μm. c, Still frames show side views of 3D volume images from time-lapse 3D SoRa SDCM imaging of GFP-SEPT9_i1 (green) and SPY650_FastAct_X (magenta) in a U2OS cell region confined in a Matrigel/laminin-filled 8 μm-wide pore. Yellow arrowheads point to short-lived actoseptin cables. White arrows point to longer lived actoseptin cables which become brighter and thicker. Blue arrows point to de novo generation of actoseptin cables that grow longitudinally and laterally. Scale bar, 1 μm.
    Pacgfp N1, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a , Top-down (top) and oblique angle (middle, bottom) views of still frames from time-lapse 3D SoRa SDCM imaging of LAP2β-AcGFP1 (green) and SPY555_FastAct_X (magenta) in the U2OS cell region confined in a Matrigel/laminin-filled 8 μm-wide pore. Images were rendered and reconstructed in 3D for each time point. Angled side views are more zoomed in on the nuclear envelope, which appears to surround the pore. Nuclear envelope creases and folds (arrows) are seen in the 60 and 120 minute time frames. Note that the nuclear envelope preserves its continuity in the 120 minute timeframe, but the front side becomes dim due to higher signal intensity in the creased regions of the 3D-rendered image. Scale bars, 2 μm. b, Time-lapse frames of a single optical slice from SoRa SDCM imaging of a U2OS cell region confined in a Matrigel/laminin-filled 8 μm-wide pore. U2OS cells were transfected with GFP-SEPT9_i1 (green) and labeled live with SPY650_FastAct_X (magents). White arrows point to actin that colocalizes with GFP-SEPT9_i1, and enlarges overtime. Blue arrows point to actoseptin that appears in frame de novo and remains spatially stable overtime. Scale bars, 2 μm. c, Still frames show side views of 3D volume images from time-lapse 3D SoRa SDCM imaging of GFP-SEPT9_i1 (green) and SPY650_FastAct_X (magenta) in a U2OS cell region confined in a Matrigel/laminin-filled 8 μm-wide pore. Yellow arrowheads point to short-lived actoseptin cables. White arrows point to longer lived actoseptin cables which become brighter and thicker. Blue arrows point to de novo generation of actoseptin cables that grow longitudinally and laterally. Scale bar, 1 μm.

    Journal: bioRxiv

    Article Title: Septins buffer actomyosin forces to protect the nucleus from genotoxic mechanical stress

    doi: 10.64898/2026.01.21.700789

    Figure Lengend Snippet: a , Top-down (top) and oblique angle (middle, bottom) views of still frames from time-lapse 3D SoRa SDCM imaging of LAP2β-AcGFP1 (green) and SPY555_FastAct_X (magenta) in the U2OS cell region confined in a Matrigel/laminin-filled 8 μm-wide pore. Images were rendered and reconstructed in 3D for each time point. Angled side views are more zoomed in on the nuclear envelope, which appears to surround the pore. Nuclear envelope creases and folds (arrows) are seen in the 60 and 120 minute time frames. Note that the nuclear envelope preserves its continuity in the 120 minute timeframe, but the front side becomes dim due to higher signal intensity in the creased regions of the 3D-rendered image. Scale bars, 2 μm. b, Time-lapse frames of a single optical slice from SoRa SDCM imaging of a U2OS cell region confined in a Matrigel/laminin-filled 8 μm-wide pore. U2OS cells were transfected with GFP-SEPT9_i1 (green) and labeled live with SPY650_FastAct_X (magents). White arrows point to actin that colocalizes with GFP-SEPT9_i1, and enlarges overtime. Blue arrows point to actoseptin that appears in frame de novo and remains spatially stable overtime. Scale bars, 2 μm. c, Still frames show side views of 3D volume images from time-lapse 3D SoRa SDCM imaging of GFP-SEPT9_i1 (green) and SPY650_FastAct_X (magenta) in a U2OS cell region confined in a Matrigel/laminin-filled 8 μm-wide pore. Yellow arrowheads point to short-lived actoseptin cables. White arrows point to longer lived actoseptin cables which become brighter and thicker. Blue arrows point to de novo generation of actoseptin cables that grow longitudinally and laterally. Scale bar, 1 μm.

    Article Snippet: Cells were transfected transiently with the plasmids pEGFP-C2_SEPT9_i1 (addgene plasmid #71609) or LAP2 Full I pAcGFP-N1 monomeric GFP (addgene plasmid #62044; gift from Dr. Eric Schirmer) using Lipofectamine 2000 transfection reagent (Invitrogen, 11668019) according to the manufacturer’s instructions with minor optimization.

    Techniques: Imaging, Transfection, Labeling