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. 2021 Mar 25;17(3):e1009422. doi: 10.1371/journal.ppat.1009422

Fig 3. NRF2 expression controls inflammation in LgyLRV1+ infected macrophages.

Fig 3

WT, Tlr3-/- and Nrf2-/-, BMDMs were infected with either LgyLRV1+ or LgyLRV1- parasites. Negative and positive controls of non-treated (Ø), tBHQ (10 μM), poly I:C (I:C, 2 μg/ml), TLR3 agonist, were performed concurrently. A) Secreted levels of TNF-α into the supernatant at 24 hr time-point by ELISA. B) Immunofluorescence of nuclear translocation of P65 subunit of NF-κB complex expressed as nucleus/cytoplasm (N/C) ratio. Cells were stained with NRF2 (red), DNA (blue) at 24 hrs and imaged at 63x using a confocal microscope. Scale bar represents 10 μm. C) Quantification of P65 nuclear translocation at 24 hrs. P65 nuclear translocation was quantified for each cell (N = 50–90) by IMARIS software. Representative images from two independent experiments are shown. Data show mean ± SEM from a pool of three (A) and two (C) independent experiments. Unpaired Student’s t test was used to assess statistical significance. Not significant (ns), * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.