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. Author manuscript; available in PMC: 2013 Apr 3.
Published in final edited form as: Cell Host Microbe. 2012 Mar 15;11(3):277–289. doi: 10.1016/j.chom.2012.01.019

Figure 3. Rubicon Inhibits CBM-Mediated Signaling in a CARD9 Binding-Dependent Manner.

Figure 3

(A) Rubicon expression suppresses β-1,3-glucan-induced downstream signaling. Raw264.7 cells containing vector, Rubicon, or its mutants were stimulated with β-1,3-glucan for the indicated times and then subjected to IB analysis to detect the phosphorylated and total forms of p38, p42/p44 MAPK, JNK, or IκB-α.

(B) Rubicon expression levels show no effect on ROS production induced by Dectin-1-mediated stimulation. BMDMs infected with lentivirus-shRNA-NS or lentivirus-shRNA-Rubicon for Rubicon depletion or infected with lenti-GFP or lenti-GFP-Rubicon for Rubicon expression were analyzed for ROS production upon stimulation with β-1,3-glucan or zymosan depleted by hot alkali treatment (to remove its TLR-stimulating properties but keep its Dectin-1-stimulating activities), or infection with SeV or VSV.

(C) Rubicon expression levels show no effect on CARD9-mediated ROS production. WT or CARD9 KO BMDMs infected with lenti-GFP or lenti-GFP-Rubicon for Rubicon expression or infected with lentivirus-shRNA-NS or lentivirus-shRNA-Rubicon for Rubicon depletion were analyzed for ROS production upon stimulation with zymosan, C. albicans, or influenza A virus infection. Values are the mean ± SD of triplicate samples. *p < 0.05 and **p < 0.01 compared with WT control cultures.

(D) Rubicon suppresses autophagosome maturation. Raw246.7 cells expressing vector, Rubicon WT, ΔSR-N, or ΔHC mutant were treated with rapamycin, starvation, β-1,3-glucan, or SeV infection for the indicated times, and their cell lysates were used for IB with αLC3, αp62, or αactin. See also Figure S3.