Transformation of the Δafoch1 mutant with the afoch4 gene (under the control of the constitutive gpdA promoter) did not rescue the sporulation phenotype on calcium plates (Supplementary Figure S4, panel C) suggesting that AfOch1 and AfOch4 have different biological functions.
Transformants expressing Ptet::clpXΔN200-dhfr1,2 and Ptet::ftsZ-dhfr3 did not grow on Trim plates (Supplementary Fig. S5, panel B) corroborating the conclusion from BACTH data that the N-terminal 200 amino acids of ClpX are needed for interaction with FtsZ.
Indeed, Trf1Δ/Δ K-Ras+ /G12V p53−/− MEFs showed a 21-fold higher percentage of senescent cells compared to Trf1+ / + K-Ras+ /G12V p53−/− MEFs 7 days post-plating (Supplementary Fig S1B), most likely reflecting an additive effect of K-Ras oncogene-induced senescence and Trf1 deficiency-induced senescence (Serrano et al, 1997; Martinez et al, 2009).
Decreased adhesion was confirmed by a colorimetric assay on fibronectin-coated plates (Supplementary Figure 5c).
Non-fluorescent (M0) cells were sorted into 24-well plates (Supplementary Material, Fig. S2A).
They appeared as one point on a normal silica gel thin layer chromatography (TLC) plate (Supplementary Figure 1a).
At E12.5, mTOR was expressed in the VZ/SVZ and the cortical plate (supplementary material Fig. S2A,B).
This procedure is amenable to automation in multiwell plates, and we obtained data from twelve 96-well plates (Supplementary file 2A, Supplementary file 2B).
By means of the pyrosequencing platform, a modified method from Roos and colleagues [ 26] was used on a 96-well plate (Supplementary Text 1).
The pro-survival Bcl-2 relatives differed in their ability to inhibit Bax toxicity when spotted onto inducing plates (Supplementary Figure 2).
We also observed that KPC cells grown on collagen were less sensitive to gemcitabine treatment than those grown on an uncoated plate (Supplementary Fig S5D).
