Review





Similar Products

99
ATCC nih 3t3 fibroblasts
In vitro cell evaluations. (a, b) Fluorescence microscopic images of <t>NIH</t> <t>3T3</t> cells stained with a live/dead kit and corresponding quantitative analysis (n = 4) (scale bars, 100 μm). (c) Cytotoxicity analysis with NIT-3T3 cells using CCK-8 kit (n = 4). (d, e) Morphological analysis of NIH 3T3 cells stained for actin (red) and nucleus (blue), with fibroblast aspect ratio analysis (scale bars, 100 μm) (n = 4). (f) Schematic illustration demonstrating the selective application of ELFS coating to the target region. (g, h) Fluorescence images showing selective adhesion of NIH 3T3 and RAW 264.7 cells to ELFS-uncoated region (n = 4) (scale bars, 100 μm). (i, j) Optical images and quantification of adhered colony-forming units (CFUs) on non-coated and ELFS-coated plates after incubation in E. coli and S. aureus suspensions for 24 h (n = 4). (k) Sequential SEM images depicting biofilm formation on non-coated and ELFS- coated stent fragments (n = 3) (scale bars, 0.5 μm). (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001). ns, not significant.
Nih 3t3 Fibroblasts, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/nih/NIH%2F3T3/pmc12924897-422-10-12
Average 99 stars, based on 1 article reviews
nih 3t3 fibroblasts - by Bioz Stars, 2026-09
99/100 stars
  Buy from Supplier

86
Bristol Myers nih small research pilot subaward
In vitro cell evaluations. (a, b) Fluorescence microscopic images of <t>NIH</t> <t>3T3</t> cells stained with a live/dead kit and corresponding quantitative analysis (n = 4) (scale bars, 100 μm). (c) Cytotoxicity analysis with NIT-3T3 cells using CCK-8 kit (n = 4). (d, e) Morphological analysis of NIH 3T3 cells stained for actin (red) and nucleus (blue), with fibroblast aspect ratio analysis (scale bars, 100 μm) (n = 4). (f) Schematic illustration demonstrating the selective application of ELFS coating to the target region. (g, h) Fluorescence images showing selective adhesion of NIH 3T3 and RAW 264.7 cells to ELFS-uncoated region (n = 4) (scale bars, 100 μm). (i, j) Optical images and quantification of adhered colony-forming units (CFUs) on non-coated and ELFS-coated plates after incubation in E. coli and S. aureus suspensions for 24 h (n = 4). (k) Sequential SEM images depicting biofilm formation on non-coated and ELFS- coated stent fragments (n = 3) (scale bars, 0.5 μm). (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001). ns, not significant.
Nih Small Research Pilot Subaward, supplied by Bristol Myers, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/nih/nih+pilot+research+small+subaward/pmc13266026-215-29-107
Average 86 stars, based on 1 article reviews
nih small research pilot subaward - by Bioz Stars, 2026-09
86/100 stars
  Buy from Supplier

99
ATCC dimethoxyethyl methacrylamide dmemam
In vitro cell evaluations. (a, b) Fluorescence microscopic images of <t>NIH</t> <t>3T3</t> cells stained with a live/dead kit and corresponding quantitative analysis (n = 4) (scale bars, 100 μm). (c) Cytotoxicity analysis with NIT-3T3 cells using CCK-8 kit (n = 4). (d, e) Morphological analysis of NIH 3T3 cells stained for actin (red) and nucleus (blue), with fibroblast aspect ratio analysis (scale bars, 100 μm) (n = 4). (f) Schematic illustration demonstrating the selective application of ELFS coating to the target region. (g, h) Fluorescence images showing selective adhesion of NIH 3T3 and RAW 264.7 cells to ELFS-uncoated region (n = 4) (scale bars, 100 μm). (i, j) Optical images and quantification of adhered colony-forming units (CFUs) on non-coated and ELFS-coated plates after incubation in E. coli and S. aureus suspensions for 24 h (n = 4). (k) Sequential SEM images depicting biofilm formation on non-coated and ELFS- coated stent fragments (n = 3) (scale bars, 0.5 μm). (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001). ns, not significant.
Dimethoxyethyl Methacrylamide Dmemam, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/nih/NIH%2F3T3/pm42319790-25-0-19
Average 99 stars, based on 1 article reviews
dimethoxyethyl methacrylamide dmemam - by Bioz Stars, 2026-09
99/100 stars
  Buy from Supplier

99
ATCC article pii s2211124725005108 nih3t3 atcc crl
In vitro cell evaluations. (a, b) Fluorescence microscopic images of <t>NIH</t> <t>3T3</t> cells stained with a live/dead kit and corresponding quantitative analysis (n = 4) (scale bars, 100 μm). (c) Cytotoxicity analysis with NIT-3T3 cells using CCK-8 kit (n = 4). (d, e) Morphological analysis of NIH 3T3 cells stained for actin (red) and nucleus (blue), with fibroblast aspect ratio analysis (scale bars, 100 μm) (n = 4). (f) Schematic illustration demonstrating the selective application of ELFS coating to the target region. (g, h) Fluorescence images showing selective adhesion of NIH 3T3 and RAW 264.7 cells to ELFS-uncoated region (n = 4) (scale bars, 100 μm). (i, j) Optical images and quantification of adhered colony-forming units (CFUs) on non-coated and ELFS-coated plates after incubation in E. coli and S. aureus suspensions for 24 h (n = 4). (k) Sequential SEM images depicting biofilm formation on non-coated and ELFS- coated stent fragments (n = 3) (scale bars, 0.5 μm). (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001). ns, not significant.
Article Pii S2211124725005108 Nih3t3 Atcc Crl, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/nih/NIH%2F3T3/pm42315647-347-21-24
Average 99 stars, based on 1 article reviews
article pii s2211124725005108 nih3t3 atcc crl - by Bioz Stars, 2026-09
99/100 stars
  Buy from Supplier

90
ATCC candida albicans
In vitro cell evaluations. (a, b) Fluorescence microscopic images of <t>NIH</t> <t>3T3</t> cells stained with a live/dead kit and corresponding quantitative analysis (n = 4) (scale bars, 100 μm). (c) Cytotoxicity analysis with NIT-3T3 cells using CCK-8 kit (n = 4). (d, e) Morphological analysis of NIH 3T3 cells stained for actin (red) and nucleus (blue), with fibroblast aspect ratio analysis (scale bars, 100 μm) (n = 4). (f) Schematic illustration demonstrating the selective application of ELFS coating to the target region. (g, h) Fluorescence images showing selective adhesion of NIH 3T3 and RAW 264.7 cells to ELFS-uncoated region (n = 4) (scale bars, 100 μm). (i, j) Optical images and quantification of adhered colony-forming units (CFUs) on non-coated and ELFS-coated plates after incubation in E. coli and S. aureus suspensions for 24 h (n = 4). (k) Sequential SEM images depicting biofilm formation on non-coated and ELFS- coated stent fragments (n = 3) (scale bars, 0.5 μm). (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001). ns, not significant.
Candida Albicans, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/nih/Genomic+DNA+from+Candida+albicans+NIH+3172/pm42320364-32-0-2
Average 90 stars, based on 1 article reviews
candida albicans - by Bioz Stars, 2026-09
90/100 stars
  Buy from Supplier

86
Wolters Kluwer Health nephrology nih guide
In vitro cell evaluations. (a, b) Fluorescence microscopic images of <t>NIH</t> <t>3T3</t> cells stained with a live/dead kit and corresponding quantitative analysis (n = 4) (scale bars, 100 μm). (c) Cytotoxicity analysis with NIT-3T3 cells using CCK-8 kit (n = 4). (d, e) Morphological analysis of NIH 3T3 cells stained for actin (red) and nucleus (blue), with fibroblast aspect ratio analysis (scale bars, 100 μm) (n = 4). (f) Schematic illustration demonstrating the selective application of ELFS coating to the target region. (g, h) Fluorescence images showing selective adhesion of NIH 3T3 and RAW 264.7 cells to ELFS-uncoated region (n = 4) (scale bars, 100 μm). (i, j) Optical images and quantification of adhered colony-forming units (CFUs) on non-coated and ELFS-coated plates after incubation in E. coli and S. aureus suspensions for 24 h (n = 4). (k) Sequential SEM images depicting biofilm formation on non-coated and ELFS- coated stent fragments (n = 3) (scale bars, 0.5 μm). (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001). ns, not significant.
Nephrology Nih Guide, supplied by Wolters Kluwer Health, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/nih/guide+nephrology+nih/10__34067_slash_kid__0000001252-13-13-2
Average 86 stars, based on 1 article reviews
nephrology nih guide - by Bioz Stars, 2026-09
86/100 stars
  Buy from Supplier

nih3t3  (ATCC)
99
ATCC nih3t3
Exploration of optogenetic clustering properties of CRY2. (A) Top panels, time-lapse images of a <t>NIH3T3</t> cell expressing CRY2high–mCherry activated with a 488-nm microscope laser starting at time t =0 (blue vertical arrow, 1.5 s pulses every 10 s). Scale bars: 5 µm. Bottom panel, coefficient of variation (CV) of fluorescence intensity calculated as the ratio between the nuclear intensity standard deviation and the nuclear intensity mean, presented as relative to the CV at time t =0. Data points corresponding to the images are marked in red. (B) Top panel, protein sequence of the C-terminus of the CRY2 PHR domain and part of the artificial linker used for C-terminal fusions for wild-type CRY2 (CRY2wt) and for CRY2 mutants. The newly generated variant CRY2hiclu is marked in bold. Mutations relative to the CRY2wt sequence are highlighted in gray. Bottom panel, images of NIH3T3 cells expressing CRY2 mutants fused to mCherry, illuminated with 1 s blue light pulses every 10 s for 15 min, and then fixed. The nucleus is delimited with a yellow line. Scale bars: 5 µm. (C) CV calculated from images obtained from NIH3T3 cells expressing CRY2 variants fused to mCherry, illuminated with pulsed blue light for 15 min, and then fixed, plotted as a function of mCherry nuclear intensity. ∼25 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple logistic fits. (D) Time-lapse images of a NIH3T3 cell expressing CRY2hiclu–mCherry activated once with the 488-nm microscope laser for 15 s at time t =0 (marked with a blue arrow). Scale bars: 5 µm. (E) Mean ( n =25) CV calculated from time-lapse images obtained from NIH3T3 cells expressing CRY2olig-mCherry, illuminated with blue light at time t =0, and then kept without blue light. The clustering ( t c ) and declustering times ( t d ) were determined from individual kinetic curves. (F) t c (top panel) and t d (bottom panel) represented as a function of mCherry nuclear intensity. ∼25–40 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple exponential (clustering) and linear (declustering) fits.
Nih3t3, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/nih/NIH%2F3T3/pmc12952762-174-0-3
Average 99 stars, based on 1 article reviews
nih3t3 - by Bioz Stars, 2026-09
99/100 stars
  Buy from Supplier

99
ATCC non tumor nih3t3 cell line
Exploration of optogenetic clustering properties of CRY2. (A) Top panels, time-lapse images of a <t>NIH3T3</t> cell expressing CRY2high–mCherry activated with a 488-nm microscope laser starting at time t =0 (blue vertical arrow, 1.5 s pulses every 10 s). Scale bars: 5 µm. Bottom panel, coefficient of variation (CV) of fluorescence intensity calculated as the ratio between the nuclear intensity standard deviation and the nuclear intensity mean, presented as relative to the CV at time t =0. Data points corresponding to the images are marked in red. (B) Top panel, protein sequence of the C-terminus of the CRY2 PHR domain and part of the artificial linker used for C-terminal fusions for wild-type CRY2 (CRY2wt) and for CRY2 mutants. The newly generated variant CRY2hiclu is marked in bold. Mutations relative to the CRY2wt sequence are highlighted in gray. Bottom panel, images of NIH3T3 cells expressing CRY2 mutants fused to mCherry, illuminated with 1 s blue light pulses every 10 s for 15 min, and then fixed. The nucleus is delimited with a yellow line. Scale bars: 5 µm. (C) CV calculated from images obtained from NIH3T3 cells expressing CRY2 variants fused to mCherry, illuminated with pulsed blue light for 15 min, and then fixed, plotted as a function of mCherry nuclear intensity. ∼25 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple logistic fits. (D) Time-lapse images of a NIH3T3 cell expressing CRY2hiclu–mCherry activated once with the 488-nm microscope laser for 15 s at time t =0 (marked with a blue arrow). Scale bars: 5 µm. (E) Mean ( n =25) CV calculated from time-lapse images obtained from NIH3T3 cells expressing CRY2olig-mCherry, illuminated with blue light at time t =0, and then kept without blue light. The clustering ( t c ) and declustering times ( t d ) were determined from individual kinetic curves. (F) t c (top panel) and t d (bottom panel) represented as a function of mCherry nuclear intensity. ∼25–40 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple exponential (clustering) and linear (declustering) fits.
Non Tumor Nih3t3 Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/nih/NIH%2F3T3/pm42280225-119-29-37
Average 99 stars, based on 1 article reviews
non tumor nih3t3 cell line - by Bioz Stars, 2026-09
99/100 stars
  Buy from Supplier

99
ATCC nih 3 t3
Exploration of optogenetic clustering properties of CRY2. (A) Top panels, time-lapse images of a <t>NIH3T3</t> cell expressing CRY2high–mCherry activated with a 488-nm microscope laser starting at time t =0 (blue vertical arrow, 1.5 s pulses every 10 s). Scale bars: 5 µm. Bottom panel, coefficient of variation (CV) of fluorescence intensity calculated as the ratio between the nuclear intensity standard deviation and the nuclear intensity mean, presented as relative to the CV at time t =0. Data points corresponding to the images are marked in red. (B) Top panel, protein sequence of the C-terminus of the CRY2 PHR domain and part of the artificial linker used for C-terminal fusions for wild-type CRY2 (CRY2wt) and for CRY2 mutants. The newly generated variant CRY2hiclu is marked in bold. Mutations relative to the CRY2wt sequence are highlighted in gray. Bottom panel, images of NIH3T3 cells expressing CRY2 mutants fused to mCherry, illuminated with 1 s blue light pulses every 10 s for 15 min, and then fixed. The nucleus is delimited with a yellow line. Scale bars: 5 µm. (C) CV calculated from images obtained from NIH3T3 cells expressing CRY2 variants fused to mCherry, illuminated with pulsed blue light for 15 min, and then fixed, plotted as a function of mCherry nuclear intensity. ∼25 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple logistic fits. (D) Time-lapse images of a NIH3T3 cell expressing CRY2hiclu–mCherry activated once with the 488-nm microscope laser for 15 s at time t =0 (marked with a blue arrow). Scale bars: 5 µm. (E) Mean ( n =25) CV calculated from time-lapse images obtained from NIH3T3 cells expressing CRY2olig-mCherry, illuminated with blue light at time t =0, and then kept without blue light. The clustering ( t c ) and declustering times ( t d ) were determined from individual kinetic curves. (F) t c (top panel) and t d (bottom panel) represented as a function of mCherry nuclear intensity. ∼25–40 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple exponential (clustering) and linear (declustering) fits.
Nih 3 T3, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/nih/NIH%2F3T3/pm42257498-317-0-4
Average 99 stars, based on 1 article reviews
nih 3 t3 - by Bioz Stars, 2026-09
99/100 stars
  Buy from Supplier

99
ATCC cell culture
Exploration of optogenetic clustering properties of CRY2. (A) Top panels, time-lapse images of a <t>NIH3T3</t> cell expressing CRY2high–mCherry activated with a 488-nm microscope laser starting at time t =0 (blue vertical arrow, 1.5 s pulses every 10 s). Scale bars: 5 µm. Bottom panel, coefficient of variation (CV) of fluorescence intensity calculated as the ratio between the nuclear intensity standard deviation and the nuclear intensity mean, presented as relative to the CV at time t =0. Data points corresponding to the images are marked in red. (B) Top panel, protein sequence of the C-terminus of the CRY2 PHR domain and part of the artificial linker used for C-terminal fusions for wild-type CRY2 (CRY2wt) and for CRY2 mutants. The newly generated variant CRY2hiclu is marked in bold. Mutations relative to the CRY2wt sequence are highlighted in gray. Bottom panel, images of NIH3T3 cells expressing CRY2 mutants fused to mCherry, illuminated with 1 s blue light pulses every 10 s for 15 min, and then fixed. The nucleus is delimited with a yellow line. Scale bars: 5 µm. (C) CV calculated from images obtained from NIH3T3 cells expressing CRY2 variants fused to mCherry, illuminated with pulsed blue light for 15 min, and then fixed, plotted as a function of mCherry nuclear intensity. ∼25 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple logistic fits. (D) Time-lapse images of a NIH3T3 cell expressing CRY2hiclu–mCherry activated once with the 488-nm microscope laser for 15 s at time t =0 (marked with a blue arrow). Scale bars: 5 µm. (E) Mean ( n =25) CV calculated from time-lapse images obtained from NIH3T3 cells expressing CRY2olig-mCherry, illuminated with blue light at time t =0, and then kept without blue light. The clustering ( t c ) and declustering times ( t d ) were determined from individual kinetic curves. (F) t c (top panel) and t d (bottom panel) represented as a function of mCherry nuclear intensity. ∼25–40 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple exponential (clustering) and linear (declustering) fits.
Cell Culture, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/nih/NIH%2F3T3/pm42280755-235-0-10
Average 99 stars, based on 1 article reviews
cell culture - by Bioz Stars, 2026-09
99/100 stars
  Buy from Supplier

Image Search Results


In vitro cell evaluations. (a, b) Fluorescence microscopic images of NIH 3T3 cells stained with a live/dead kit and corresponding quantitative analysis (n = 4) (scale bars, 100 μm). (c) Cytotoxicity analysis with NIT-3T3 cells using CCK-8 kit (n = 4). (d, e) Morphological analysis of NIH 3T3 cells stained for actin (red) and nucleus (blue), with fibroblast aspect ratio analysis (scale bars, 100 μm) (n = 4). (f) Schematic illustration demonstrating the selective application of ELFS coating to the target region. (g, h) Fluorescence images showing selective adhesion of NIH 3T3 and RAW 264.7 cells to ELFS-uncoated region (n = 4) (scale bars, 100 μm). (i, j) Optical images and quantification of adhered colony-forming units (CFUs) on non-coated and ELFS-coated plates after incubation in E. coli and S. aureus suspensions for 24 h (n = 4). (k) Sequential SEM images depicting biofilm formation on non-coated and ELFS- coated stent fragments (n = 3) (scale bars, 0.5 μm). (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001). ns, not significant.

Journal: Bioactive Materials

Article Title: Slippery dopamine–fluoropolymer hybrid surface for improving biliary stent longevity

doi: 10.1016/j.bioactmat.2026.02.003

Figure Lengend Snippet: In vitro cell evaluations. (a, b) Fluorescence microscopic images of NIH 3T3 cells stained with a live/dead kit and corresponding quantitative analysis (n = 4) (scale bars, 100 μm). (c) Cytotoxicity analysis with NIT-3T3 cells using CCK-8 kit (n = 4). (d, e) Morphological analysis of NIH 3T3 cells stained for actin (red) and nucleus (blue), with fibroblast aspect ratio analysis (scale bars, 100 μm) (n = 4). (f) Schematic illustration demonstrating the selective application of ELFS coating to the target region. (g, h) Fluorescence images showing selective adhesion of NIH 3T3 and RAW 264.7 cells to ELFS-uncoated region (n = 4) (scale bars, 100 μm). (i, j) Optical images and quantification of adhered colony-forming units (CFUs) on non-coated and ELFS-coated plates after incubation in E. coli and S. aureus suspensions for 24 h (n = 4). (k) Sequential SEM images depicting biofilm formation on non-coated and ELFS- coated stent fragments (n = 3) (scale bars, 0.5 μm). (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001). ns, not significant.

Article Snippet: The prepared stents were placed on the Transwell insert, and NIH-3T3 fibroblasts (ATCC CRL-1658; 0.5 × 10 5 cells mL −1 ) or human biliary epithelial SNU-1079 cells (Korean Cell Line Bank, KCLB No. 01079; 0.5 × 10 5 cells mL −1 ) were cultured in 2 mL of DMEM supplemented with 10% bovine calf serum and 1% penicillin–streptomycin.

Techniques: In Vitro, Fluorescence, Staining, CCK-8 Assay, Incubation

Exploration of optogenetic clustering properties of CRY2. (A) Top panels, time-lapse images of a NIH3T3 cell expressing CRY2high–mCherry activated with a 488-nm microscope laser starting at time t =0 (blue vertical arrow, 1.5 s pulses every 10 s). Scale bars: 5 µm. Bottom panel, coefficient of variation (CV) of fluorescence intensity calculated as the ratio between the nuclear intensity standard deviation and the nuclear intensity mean, presented as relative to the CV at time t =0. Data points corresponding to the images are marked in red. (B) Top panel, protein sequence of the C-terminus of the CRY2 PHR domain and part of the artificial linker used for C-terminal fusions for wild-type CRY2 (CRY2wt) and for CRY2 mutants. The newly generated variant CRY2hiclu is marked in bold. Mutations relative to the CRY2wt sequence are highlighted in gray. Bottom panel, images of NIH3T3 cells expressing CRY2 mutants fused to mCherry, illuminated with 1 s blue light pulses every 10 s for 15 min, and then fixed. The nucleus is delimited with a yellow line. Scale bars: 5 µm. (C) CV calculated from images obtained from NIH3T3 cells expressing CRY2 variants fused to mCherry, illuminated with pulsed blue light for 15 min, and then fixed, plotted as a function of mCherry nuclear intensity. ∼25 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple logistic fits. (D) Time-lapse images of a NIH3T3 cell expressing CRY2hiclu–mCherry activated once with the 488-nm microscope laser for 15 s at time t =0 (marked with a blue arrow). Scale bars: 5 µm. (E) Mean ( n =25) CV calculated from time-lapse images obtained from NIH3T3 cells expressing CRY2olig-mCherry, illuminated with blue light at time t =0, and then kept without blue light. The clustering ( t c ) and declustering times ( t d ) were determined from individual kinetic curves. (F) t c (top panel) and t d (bottom panel) represented as a function of mCherry nuclear intensity. ∼25–40 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple exponential (clustering) and linear (declustering) fits.

Journal: Journal of Cell Science

Article Title: OptoLoop – an optogenetic tool to probe the functional role of genome organization

doi: 10.1242/jcs.264574

Figure Lengend Snippet: Exploration of optogenetic clustering properties of CRY2. (A) Top panels, time-lapse images of a NIH3T3 cell expressing CRY2high–mCherry activated with a 488-nm microscope laser starting at time t =0 (blue vertical arrow, 1.5 s pulses every 10 s). Scale bars: 5 µm. Bottom panel, coefficient of variation (CV) of fluorescence intensity calculated as the ratio between the nuclear intensity standard deviation and the nuclear intensity mean, presented as relative to the CV at time t =0. Data points corresponding to the images are marked in red. (B) Top panel, protein sequence of the C-terminus of the CRY2 PHR domain and part of the artificial linker used for C-terminal fusions for wild-type CRY2 (CRY2wt) and for CRY2 mutants. The newly generated variant CRY2hiclu is marked in bold. Mutations relative to the CRY2wt sequence are highlighted in gray. Bottom panel, images of NIH3T3 cells expressing CRY2 mutants fused to mCherry, illuminated with 1 s blue light pulses every 10 s for 15 min, and then fixed. The nucleus is delimited with a yellow line. Scale bars: 5 µm. (C) CV calculated from images obtained from NIH3T3 cells expressing CRY2 variants fused to mCherry, illuminated with pulsed blue light for 15 min, and then fixed, plotted as a function of mCherry nuclear intensity. ∼25 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple logistic fits. (D) Time-lapse images of a NIH3T3 cell expressing CRY2hiclu–mCherry activated once with the 488-nm microscope laser for 15 s at time t =0 (marked with a blue arrow). Scale bars: 5 µm. (E) Mean ( n =25) CV calculated from time-lapse images obtained from NIH3T3 cells expressing CRY2olig-mCherry, illuminated with blue light at time t =0, and then kept without blue light. The clustering ( t c ) and declustering times ( t d ) were determined from individual kinetic curves. (F) t c (top panel) and t d (bottom panel) represented as a function of mCherry nuclear intensity. ∼25–40 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple exponential (clustering) and linear (declustering) fits.

Article Snippet: NIH3T3 (mouse fibroblasts, ATCC #CRL-1658), U2OS (from human osteosarcoma, ATCC #HTB-96), HeLa (from human cervical adenocarcinoma, ATCC #CRM-CCL-2) and Lenti-X HEK-293T (from human embryonic kidney, cat. #632180 from Takara Bio, Japan) cell lines were cultured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) supplemented with 10% (15% for NIH3T3) fetal bovine serum (Gibco, Waltham, MA, USA) plus 100 IU/ml penicillin and 100 μg/ml streptomycin (Gibco, Waltham, MA, USA) at 37°C in a humidified atmosphere with 5% CO 2 .

Techniques: Expressing, Microscopy, Fluorescence, Standard Deviation, Sequencing, Generated, Variant Assay