let’s do the full summary later
here is the recent summary for the 2024 committee meeting
after the previous work finishing in mid 2022, we have been trying to study how Piezo1 regulate its location inside the cell. we found that in tdTMEF cells, when they overlap on the edges, Piezo1 signal shows a 5 fold difference over reference membrane markers.
put a file link here
this leads to a thinking of cell-cell adhesion could play a role in regulating Piezo1 activity.
since Sajad has the beautiful L929 cell line to work on cell-cell adhesion in suspension culture, we decided to try the L929 cell line first. which didn’t quite work out eventually. Mainly due to
- low expression level of Piezo1
- hard to make them attach to each other
- wide field fluorescence imaging is not capable enough to show the difference on the suspension(too many scattering?)
so we decided to move on to some adherent cell culture. which looks equally hard.
Piezo1 and Piezo2 comparison
aside from cell-cell adhesion project. because we got some Piezo2-mScarlet protein from stefan G. lechner we also tried to work on those to see if there is any localization difference between these homologs.
Updates0508 P1 and P2 colocalization analysis P2 bleb pulling raw PTK2 cells
Updates0516 PTK2-tubulin cells with 14 79 transfection intracellular bridge
Updates0529 14 79 83 long term imaging 38 116 79 tether pulling
Updates0605 38 79 116 poking P1 P2 colocalization analysis
Updates0612 P1 P2 latB blebs
Updates0626 tdTMEF suspension tdTMEF TIRF imaging test PK640 dye and deep red dye, both shows strong bleachthrough in red channel.
Updates0717 NEM treated P1 P2 GPMVs (P2 not showing membrane existence) P1 P2 10X imaging P1 P2 yoda1
later on, it is the bead rotation and GPMV aspiration experiment which is familiar to me now. let’s work on that later.