Cell growth experiments were performed by plating ARPE-19 cells on 12-well plates (Sigma-Aldrich, St . Louis MO, USA) at a concentration of 200,000 cells/ml/well in either serum-free or serum-containing medium.
Untreated cells or cells treated with dimethyl sulfoxide (DMSO, Sigma-Aldrich, St . Louis MO, USA) at a concentration identical to that present in the fisetin or luteolin samples served as negative controls (0.25% v/v).
Resveratrol samples were dissolved in dimethyl sulfoxide (DMSO) (Sigma, USA) at a concentration of 200 mM, and diluted in culture medium just before use.
Fresh diets were prepared daily by adding MRS broth (Merck) to the pre-mixture (Research Diets Inc., New Brunswick, NJ, USA) at a concentration of 20% (v/w).
For orthotopic inoculation, the cells were trypsinised and suspended in Matrigel (BD Biosciences, San Jose, CA, USA) at a concentration of 106 cells/mL and stored on ice.
The membrane was incubated for 1 h at room temperature with mouse anti-FLAG M2-Peroxidase antibody (Sigma-Aldrich, St Louis, MO, USA) at a concentration of 1 10000 in 5% skim milk/PBS.
The cleavage reaction for the fusion protein was performed using the enzyme enterokinase EKMax (Invitrogen, USA) at a concentration of 1 U per 20 μg of purified recombinant peptide and incubated at 22°C overnight.
The cells were incubated with LPS (Escherichia coli 0111 B4, Sigma-Aldrich, Inc. USA) at a concentration of 0.625 µg/ml for 24 hours [22].
The protein loads on the blot were checked by staining with anti-ERK2 total antibody (item K-23 #sc-153, Santa Cruz Biotechnology, Santa Cruz, CA, USA) at a concentration suggested by the supplier.
M. bovis BCG Pasteur 1173P2 was grown to mid log phase in Souton's medium and treated with D- Mannosamine (ManN) (Sigma Aldrich, USA) at a concentration of 5 mg/ml.
RINm5F cells were cultured in chambered slides, fixed with 4% formaldehyde in PBS and incubated with antibodies to OGT (Santa Cruz Biotechnology, USA), and PHB (Cell Signaling, USA) at a concentration of 1 µg/ml in PBS containing 0.1% BSA.
