10/24/23
10/24/23 kdorfman Mon, 10/16/2023 - 19:36Students begin projects
Kate provides fixed coverslips (seeded 10/22, fixed 10/24/23)
| Cell line | medium | variant | treatment | no. coverslips |
|---|---|---|---|---|
| HCT116 | McCoy | WT (60) | 4 | |
| LB1-AID-GFP (68) | 2 | |||
| MEF | DMEM | WT (39) | control | 6 |
| VPA | ||||
| DZNP | ||||
| LMNB1-/- (02) | 3 | |||
| DMEM+G418 | LA KD (37) | 3 | ||
| Prostate cancer | RPMI-1640 | DU145 (74) | 2 +3 from Mai | |
| RPMI-1640 | LNCaP (77) | 2 + 3 from Mai | ||
| DMEM | PC3 (76) | 2 |
Materials:
- Paraglut fix (enough for 30 wells)
- PBS to rinse 30 coverslips
- PB-TW-Azide (enough for 24 dunking beakers and 30 wells)
- 1o antibodies
- mouse anti-NP (nuclear pore)
- mouse anti-LB1
- rabbit anti-LB2 ab151735 (get one 10 µL aliquot from Drew - dilute 1/1000)
- 2o antibodies
- goat anti-mouse green for NP
- goat anti-rabbit red for LB1 & LB2
- Slides
- slide labels
- DAPI & droppers
- forceps
- dunking beakers
- humid chamber materials
| Project | people | scope | IF coverslips | IF details |
|---|---|---|---|---|
| MEF VPA | Olivia & Juliana | 1 | 1x 39 unt, 1X 39 VPA, another VPA if pos | standard NP LB1 IF |
| MEF DZNep | Karan & Nate | 2 | 1x 39 unt, 1X 39 DZNep, another if pos | standard NP LB1 IF |
| MEF LMNB1-/- | Cole & Grif | 3 | 1X 39, 2X 02 LMNB1-/- | 39 & 02 NP LB2 IF, 1x 02 NP LB1 IF |
| MEF LA KD | Shrushti, Emma, Anna | 8 | 1X 39, 2X 39 | standard NP LB1 IF |
| HCT116 LB1 live 1 | Sam & Jillian | 5 | 68, 1x unt & 1x VPA | LB2 IF only |
| HCT116 LB1 live 2 | Isabelle & Jason | 6 | 60, 1x unt 1x VPA; 60, 1x unt 1x VPA | one set NP LB1 IF; other set NP LB2 IF |
| Prostate Cancer 1 | Tyian & Antonia | 4 | 2x 74, 1x 76 | standard NP LB1 IF |
| Prostate Cancer 2 | Kelsey & Pedro | 7 | 2x 74, 1x 77 | standard NP LB1 IF |