Cells were incubated with Lipofectamine and DNA (3:1 ratio) for 5 h in growth medium devoid of all supplements, after which time medium was changed to complete growth medium.
Liquid cultures were performed in shaker flasks (220 rpm; 30°C) in R5− medium (R5 medium devoid of additional K2HPO4, CaCl2 and L-proline) as described earlier [32].
While M. smegmatis grew normally in medium devoid of platensimycin, the culture in the medium containing platensimycin showed a decrease in OD600 values with time (data not shown) resulting in clumping after 24 hours of incubation (Figure 3A).
Four kinds of treatment of the cells were tested: (a) in normal cell culture medium, as described above, (b) in cell culture medium devoid of fetal calf serum, (c) in cell culture medium supplemented with 1 µM gefitinib (Biaffin GmbH, Kassel, Germany) and (d) in cell culture medium devoid of serum but with 1 µM gefitinib.
To validate our in silico data and test whether autophagy could be induced by nutrient limitation in P. falciparum, parasite cultures were grown in both serum-free medium and minimal medium devoid of the essential amino acid L-glutamine and serum, with minimal glucose.
On the next day, the volume of incubation medium was diluted (1 : 1) with the addition of medium devoid of FBS.
The growth medium was replaced with DCSS medium devoid of antimycotic compound and cultured for an additional 3 days.
Fully polarized MDCKII cell monolayers were switched from a medium containing calcium to a medium devoid of calcium to induce junctional complex disassembly (Ivanov et al, 2004 and see Section 4).
They were either stimulated in this medium (SM10) by the addition of 30 μg/ml of heat inactivated E. coli or S. aureus particles for various periods of time, or the cultures were washed twice with PBS and the culture medium was replaced by growth medium devoid of FCS (SM0) and the cells stimulated similarly as in SM10.
Mn-treated larvae (48-hour exposure to MnCl2) were washed several times with normal E3 medium and transferred back to E3 medium devoid of Mn.
Adk−/− or wild type (wt) ESC-derived glial precursor cells were seeded on the substrates and cultured either in proliferation medium containing growth factors or in differentiation medium devoid of growth factors.
