Journal: Communications Biology

Article Title: Optogenetic control of integrin-matrix interaction

doi: 10.1038/s42003-018-0264-7

Figure Lengend Snippet: Validation of cell–matrix interaction with different cell lines stably expressing OptoIntegrin. a HEK-293T, HeLa and MCF7 cells stably expressing OptoIntegrin or moxGFP-PIF S were seeded on PhyB 1–651 -coated glass slides and incubated under 660 nm or 740 nm light (I = 20 µmol m −2 s −1 ) for 5 h and subsequently imaged using a transmission light microscope (scale bar = 200 µm). b Live cell imaging of cell–matrix interaction under 660 nm light and then switch to 740 nm light after 35 min with binary images of cell shapes to illustrate the cells spreading. For this, HeLa cells stably expressing OptoIntegrin were seeded on OptoMatrix and subsequently imaged. Micrographs were taken at indicated time points (scale bar = 10 µm). c Spatial control of cell attachment with OptoIntegrin-expressing HEK-293T cells. OptoIntegrin-expressing cells were cultivated on OptoMatrix, locally illuminated with 660 nm or 740 nm for 3 min and then left in darkness for 4 h. Afterwards, cells were fixed and imaged (scale bar = 200 µm)

Article Snippet: Finally, the stained coverslips were mounted on microscope slides with Mowiol 4-88 (Carl Roth, cat. no.: 0713), and confocal images were acquired on an upright confocal microscope (Nikon Instruments Eclipse Ni-E with a C2 confocal laser scanner, 100 × oil objective NA = 1.45 or 60× oil objective NA = 1.40).

Techniques: Biomarker Discovery, Stable Transfection, Expressing, Incubation, Transmission Assay, Light Microscopy, Live Cell Imaging, Control, Cell Attachment Assay

Journal: Oncotarget

Article Title: Drug-induced ciliogenesis in pancreatic cancer cells is facilitated by the secreted ATP-purinergic receptor signaling pathway

doi: 10.18632/oncotarget.23335

Figure Lengend Snippet: ( A ) Quantification of the effect of apyrase treatment on ciliogenesis. ( B ) Representative images showing the effect of apyrase on ciliogenesis of cells exposed to the indicated drugs. Nuclei were stained with DAPI (blue) and cilia with an antibody against the cilium marker acetylated tubulin (green). All images were captured using Nikon C2 Eclipse Ni-E confocal microscope using a 60× objective lens. Data are presented as mean ± SEM, * p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.0005. ( C ) Representative images showing the staining of cilia with an antibody against IFT88 (red), an alternative marker of the cilium.

Article Snippet: Images of primary cilia were captured by acquiring Z-stacks using Olympus FluoView-FV 1000 or Nikon C2 Eclipse Ni-E confocal laser scanning microscope using 60X oil immersion lens.

Techniques: Staining, Marker, Microscopy

Journal: Oncotarget

Article Title: Drug-induced ciliogenesis in pancreatic cancer cells is facilitated by the secreted ATP-purinergic receptor signaling pathway

doi: 10.18632/oncotarget.23335

Figure Lengend Snippet: ( A ) Quantification of extracellular ATP in cells treated with pannexin channel blocker (mefloquine), vesicular transport inhibitor (latrunculin B). ( B ) Quantification of the effect of mefloquine on ciliogenesis in CFPAC-1 cells. ( C ) Representative images showing the effect of pannexin channel blockade on ciliogenesis in CFPAC-1 cells exposed to a selection of ciliogenic drugs. Nuclei were stained with DAPI (blue) and cilia with an antibody against the cilium marker acetylated tubulin (green). All images were captured using Nikon C2 Eclipse Ni-E confocal microscope using a 60× objective lens. Data are presented as mean ± SEM, * p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.0005.

Article Snippet: Images of primary cilia were captured by acquiring Z-stacks using Olympus FluoView-FV 1000 or Nikon C2 Eclipse Ni-E confocal laser scanning microscope using 60X oil immersion lens.

Techniques: Selection, Staining, Marker, Microscopy

Journal: Oncotarget

Article Title: Drug-induced ciliogenesis in pancreatic cancer cells is facilitated by the secreted ATP-purinergic receptor signaling pathway

doi: 10.18632/oncotarget.23335

Figure Lengend Snippet: ( A ) Effect of suramin, a blocker of P2 purinergic receptors on ciliogenesis in CFPAC-1 cells exposed to ciliogenic drugs. ( B ) Representative images of CFPAC-1 cells showing the effect of suramin on ciliation in untreated and treated cells. Nuclei were stained with DAPI (blue) and cilia with an antibody against the cilium marker acetylated tubulin (green). All images were captured using Nikon C2 Eclipse Ni-E confocal microscope using a 60× objective lens. Data are presented as mean ± SEM, * p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.0005.

Article Snippet: Images of primary cilia were captured by acquiring Z-stacks using Olympus FluoView-FV 1000 or Nikon C2 Eclipse Ni-E confocal laser scanning microscope using 60X oil immersion lens.

Techniques: Staining, Marker, Microscopy