Identification of a Lysosomal Pathway That Modulates Glucocorticoid Signaling and the Inflammatory Response

Yuanzheng He,* Yong Xu, Chenghai Zhang, Xiang Gao, Karl J. Dykema, Katie R. Martin, Jiyuan Ke, Eric A. Hudson, Sok Kean Khoo, James H. Resau, Arthur S. Alberts, Jeffrey P. MacKeigan, Kyle A. Furge, H. Eric Xu*

*To whom correspondence should be addressed. E-mail: eric.xu{at}vai.org (H.E.X.); ajian.he{at}vai.org (Y.H.)

This PDF file includes:

• Fig. S1. Activity of CQ and amodiaquine (AQ).
• Fig. S2. Effect of CQ and bafilomycin A1 on GR localization in AD293 cells.
• Fig. S3. CQ enhances glucocorticoid-mediated GR signaling.
• Fig. S4. Microarray analysis of the effect of CQ/Dex on gene expression in THP-1 cells.
• Fig. S5. CQ inhibition of lysosomes leads to accumulation of autophagosomes and autolysosomes.
• Fig. S6. Knockdown efficiency of the siRNAs targeting components of the VATPase.
• Fig. S7. CQ stabilizes GR in AD293 cells.
• Fig. S8. The effect of CQ on a β-catenin–activated reporter gene.
• Fig. S9. Effect of the proteasome inhibitor MG132 and the lysosomal inhibitor CQ on GR activation in AD293 cells and COS7 cells.
• Table S1. Top 70 genes regulated by Dex in LPS-stimulated THP-1 cells.
• Table S2. Top 70 genes regulated by CQ in LPS-stimulated THP-1 cells.
• Table S3. Top 70 genes regulated by Dex + CQ in LPS-stimulated THP-1 cells.
• Table S4. Top 70 genes showing a synergistic transactivation effect upon Dex + CQ treatment in LPS-stimulated THP-1 cells.
• Table S5. Top 70 genes showing a synergistic transrepression effect upon Dex + CQ treatment in LPS-stimulated THP-1 cells.
• Table S6. Real-time PCR primers for target genes.
• Table S7. siRNA target sequences.
• Descriptions for Videos S1 to S3
  

[Download PDF]

Technical Details

Format: Adobe Acrobat PDF

Size: 1.60 MB

Other Supplementary Material for this manuscript includes the following:

• Video S1 (.avi format). Pulse chase of Halo-GR in AD293 cells with PBS vehicle.
• Video S2 (.avi format). Pulse chase of Halo-GR in AD293 cells exposed to CQ.
• Video S3 (.avi format). Dynamic association of GR and lysosomes in AD293 cells.

[Download Videos S1 to S3 (Compressed)]

Technical Details

Format: AVI

Sizes: 56.9 MB (compressed);
63.8 MB (Video S1 decompressed); 63.8 MB (Video S2 decompressed); 122.6 MB (Video S3 decompressed)

Instructions for downloading and decompressing files:

1. Create a temporary folder on your machine's hard drive.
2. Save the compressed archive to the temporary folder you created, using the links above.
3. Decompress the compressed file in the temporary folder using decompression software such as WinZip (Windows; www.winzip.com) or StuffIt Expander (Windows and Mac; www.stuffit.com).

© 2011 American Association for the Advancement of Science