In the article “Aberrant TGFβ signaling reduces T regulatory cells in ICAM-1 deficient mice, increasing the inflammatory response to Mycobacterium tuberculosis,” by Hillarie Plessner Windish, P. Ling Lin, Joshua T. Mattilla, Angela M. Green, Ezenwa Obi Onuoha, Lawrence P. Kane and JoAnne L. Flynn, which appeared in the Journal of Leukocyte Biology. September 2009 86:713–725; doi: 10.1189/jlb.1208740, one of the representative flow cytometry dot plots (WT-WT) in Figure 2 was inadvertently duplicated. The revised figure and figure legend appear below:
Figure 2. ICAM-1 expression is not required for migration of T regulatory cells in the periphery.
The in vivo migration of T regulatory cells from M. tuberculosis-infected mice stained red (ICAM-1–/–) or green (WT) in M. tuberculosis-infected recipients following adoptive transfer by intravenous injection. (A) Representative dot plots: detection of ICAM-1–/– (PKH26, red) or WT (PKH67, green) transferred cells in the lungs of recipient mice by flow cytometry. The dot plots shown here were gated on the lymphocytes within the live cell gates. (B) To determine whether migration requires ICAM-1 on the endothelium, the percentages of injected WT T regulatory cells detected in the lungs of ICAM-1–/– (squares) or WT (triangles) recipients (cells detected in organs/injected cells · 100) were compared. No significant differences were observed. (C) To determine whether migration requires ICAM-1 on the T regulatory cells, the percentages of transferred WT (triangles) or ICAM-1–/– (squares) T regulatory cells detected in the lungs of WT mice were compared (cells detected in organs/injected cells · 100). No significant differences were found.
The methods for stripping the Western blot shown in Figure 4 were also omitted, and Supplemental Methods and updated figure legend have been added and appear below:
Figure 4. ICAM-1 expression is not required for the association of pSmad3 and Smad4. Naïve CD4+CD25+ spleen cells from WT or ICAM1–/– mice were incubated with anti-CD3, IL-2, and TGF-β1 to generate inducible T regulatory cells, identified by their expression of FoxP3. Smad4 was immunoprecipitated from nuclear and cellular lysates, and coimmunoprecipitation was assessed by immuoblotting. (A) Representative immunoblots of coimmunoprecipitated Smad4 and pSmad3. Immunoblots were first probed for Smad4, imaged, and stripped. After confirming that no antibody for Smad4 remained, stripped blots were reprobed for pSmad3. Molecular weights shown for pSmad3 and Smad 4 reflect the known molecular weights for these proteins, not the molecular weights that the coimmunoprecipiated proteins reached. (B) Mean intensity of the bands detected in immunoblots from nuclear lysates of ICAM1–/– (Δ) and WT (○) T cells. (C) Mean intensity of the bands detected in immunoblots from cytoplasmic lysates of ICAM1–/– (Δ) and WT (○) T cells. (D) Ratio of pSmad3:Smad4 bands of nuclear and cytoplasmic lysates from ICAM1–/– (Δ) and WT (○) T cells.
Supplemental methods
Stripping immunoblot for reblotting.
After initial blot staining was imaged, immunoblots were stripped using Restore Western Blot Stripping Buffer (Thermo Scientific, Rockford, IL, USA) to each blot for 5–15 min under gentle agitation. Blots were washed and developed using chemiluminescent substrate to confirm that antibody from the first blot staining was fully removed. Immunoblots were reblocked and stained as described above.
This has been corrected online, and the authors regret any inconvenience caused by this error.
