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. 2018 May 7;217(5):1739–1755. doi: 10.1083/jcb.201709137

Figure 6.

Figure 6.

LPS-stimulated S729A B cells fail to respond to secondary ER stress by producing additional XBP1s. (A) 2-d LPS-stimulated WT and S729A B cells were exposed to 20 µM B-I09 or 5 mM DTT for 3 h, stained for XBP1s and B220, and gated for B220+/XBP1s+ populations. (B) 2-d LPS-stimulated WT and S729A B cells were exposed to 2.5 µM Tg for the indicated times, stained for XBP1s and B220, and gated for B220+/XBP1s+ populations. (C) 2-d LPS-stimulated WT and S729A B cells were exposed to 1.5 nM SubAB for the indicated times, stained for XBP1s and B220, and gated for B220+/XBP1s+ populations. (D) 2-d LPS-stimulated WT and S729A B cells were exposed to 1 nM SubAB, 5 µg/ml Tu, or 2.5 µM Tg for 12 h or were exposed to 5 mM DTT for 3 h and immunoblotted. (E) 2-d LPS-stimulated WT and S729A B cells were exposed to 1 nM SubAB or 2.5 µM Tg for 12 h or were exposed to 5 mM DTT for 3 h, lysed, and immunoprecipitated for IRE1. Bead-bound IRE1 was treated with CIP for 3 h and immunoblotted for IRE1. Data in this figure are representative of three independent experiments.