Activation of SKN-1 does not require IRE-1-TRAF (<i>trf-1</i>), when exposed to the <i>S</i>. <i>gordonii</i>.

(A) Representative images of the localization of SKN-1B/C::GFP in ire-1, trf-1 knockdown and vector control treated worms on S. gordonii for 2 hours. (B) The degree of SKN-1B/C::GFP nuclear localization and the percentage of worms in each category fed on ire-1, trf-1 knockdown and vector control treated worms on S. gordonii. A total of more than 100 worms exposed to each strain were imaged and the experiment was repeated 3 times. Similar levels of nuclear localization of SKN-1B/C::GFP was observed in the ire-1, trf-1 knockdown and vector control treated worms on S. gordonii. (C) Representative images of gcs-1::gfp expression in nsy-1, sek-1, pmk-1, skn-1 knockdown and vector control treated worms on S. gordonii. (D) The level of gcs-1::gfp expression and the percentage of worms in each category fed on ire-1, trf-1 knockdown and vector control treated worms on S. gordonii for 2 hours. A total of more than 100 worms exposed to each strain were imaged and the experiment was repeated 3 times. No difference of gcs-1::gfp expression was observed in the ire-1, trf-1 knockdown worms compared to the vector control treated worms on S. gordonii. (E) Western blot was used to analyze the level of phosphorylation of PMK-1 in vector control treated worms, ire-1 mutant worms and trf-1 knockdown worms exposed to S. gordonii for 2 hours. No significant differences was observed in the level of phosphorylation of PMK-1 in the ire-1 mutant worms and trf-1 knockdown worms compared to the vector control treated worms. α-tubulin was used as a loading control. The experiment was performed three times.