Svelty

To establish the fermentation conditions, as culture media, whole milk (with lactose) and non-lactose milk of commercial origin, both with low fat content (0.6–1.13%), were used, both set as control groups and identified as low-fat whole milk (SM) (Grupo Lala, S.A.B. de C.V., Durango, México) and lactose-free milk (LFM) (Svelty, Nestlé México S.A de C.V.). After establishing the conditions for optimizing the fermentation process, a second fermentation was carried out using additional sources of carbon to the lactose-free milk.
As additional carbon sources, there were added monosaccharides: anhydrous dextrose (27740, Golden Bell, Querétaro, Mexico), fructose (USP, 08431, Fermont, Monterrey, Mexico) and galactose (G0750, Sigma-Aldrich, St. Louis, MO, USA); the disaccharide lactose (1.07657.1000, Millipore, Burlington, MA, USA); and fructooligosaccharides: chicory inulin (Beneo^TM ST, Orafti, Mexico) and agave fructans, from Agave angustifolia Haw., obtained at CEPROBI-IPN (patent 380041). As biological material, milk kefir grains from Teziutlán, Puebla province, were utilized.

Cells were washed with cold PBS and lysed using NP40 (SIGMA-Aldrich, St. Louis, MO, USA). Lysates were centrifuged at 10,000 rpm for 10 min at 4 °C. Proteins were separated on SDS-PAGE polyacrylamide by molecular weight and transferred to PVDF membranes. Transferred membranes were blocked with 5% low-fat milk (Svelty, Nestlé) in TBS-T. Membranes were incubated overnight at 4 °C with primary antibodies p-Akt1/2 and total akt1/2 (Cell Signaling, Danvers, MA, USA) as described previously in [41 (link)].