My Beatiful Western Blot
This is where my weekend went.
The groups of bands in order from top to bottom are: ICP0, early viral protein (HSV-1, Herpes Simplex 1); beta Actin, control - just a regular cell housekeeping protein; US11, late viral protein (HSV-1). From right to left, the wells were: M-V, M-FL, M-56, M-CoT, HSV-1-V, HSV-1-FL, HSV-1-56, HSV-1-CoT, +IFN-V, +IFN-FL, +IFN-56, +IFN-CoT. M = mock (not infected with HSV-1, so accordingly, there's no viral protein). HSV-1 means infected with HSV-1. +IFN means pretreated with IFN (interferon, gamma and alpha). V = vector, no pml transfected. FL = full length, transfected with full length nuclear PML. -56 = deleted 5, 6, so transfected with the cytoplasmic form that doesn't have exons five and six. CoT = cotransfection, transfected with both FL and -56. PML is the promyelocytic leukemia protein. It has an IFN receptor on its promoter region, so people think it has something to do with anti-viral response since IFN is widely accepted to correlate with anti-viral response.
A lighter exposure. See how the US11 and ICP0's darker with FL PML versus -56? That means the infection is worse. ^_______^
That was supposed to be PML... but we don't know what happened to it. Nuclear PML usually shows up around where those really dark bands are... but the vector definitely weren't transfected with PML, and some have the cytoplasmic form which are supposed to be just a little below the nuclear form, like around 64kb. And these are mouse PML -/- fibroblasts... so definitely shouldn't have been anything... We were trying out a new antibody... guess it doesn't work like we'd hoped.
The groups of bands in order from top to bottom are: ICP0, early viral protein (HSV-1, Herpes Simplex 1); beta Actin, control - just a regular cell housekeeping protein; US11, late viral protein (HSV-1). From right to left, the wells were: M-V, M-FL, M-56, M-CoT, HSV-1-V, HSV-1-FL, HSV-1-56, HSV-1-CoT, +IFN-V, +IFN-FL, +IFN-56, +IFN-CoT. M = mock (not infected with HSV-1, so accordingly, there's no viral protein). HSV-1 means infected with HSV-1. +IFN means pretreated with IFN (interferon, gamma and alpha). V = vector, no pml transfected. FL = full length, transfected with full length nuclear PML. -56 = deleted 5, 6, so transfected with the cytoplasmic form that doesn't have exons five and six. CoT = cotransfection, transfected with both FL and -56. PML is the promyelocytic leukemia protein. It has an IFN receptor on its promoter region, so people think it has something to do with anti-viral response since IFN is widely accepted to correlate with anti-viral response.
A lighter exposure. See how the US11 and ICP0's darker with FL PML versus -56? That means the infection is worse. ^_______^
That was supposed to be PML... but we don't know what happened to it. Nuclear PML usually shows up around where those really dark bands are... but the vector definitely weren't transfected with PML, and some have the cytoplasmic form which are supposed to be just a little below the nuclear form, like around 64kb. And these are mouse PML -/- fibroblasts... so definitely shouldn't have been anything... We were trying out a new antibody... guess it doesn't work like we'd hoped.