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. 2015 Jan 20;4:e04494. doi: 10.7554/eLife.04494

Figure 1. HOIL-1 KO mice are highly susceptible to acute infection with Listeria monocytogenes, Toxoplasma gondii and Citrobacter rodentium.

(A) Survival of control (blue circles) and HOIL-1 KO (red squares) mice following i.p. inoculation with 105 (left panel; control n = 35, HOIL-1 KO n = 19), 104 (middle panel; control n = 15, HOIL-1 KO n = 15) or 103 (right panel; control n = 15, HOIL-1 KO n = 15) CFU Listeria strain EGD. (B) Listeria CFU in spleen and liver from control (blue circles) and HOIL-1 KO (red squares) mice infected with 105 CFU i.p. for 1 day (left panel), 3 days (middle panel) or 6 days (right panel). Each symbol represents an individual mouse and the mean log10 CFU is indicated. The dashed line indicates the limit of detection. (C) Survival of control (blue circles) and HOIL-1 KO (red squares) mice following inoculation with 5000 (left panel; control n = 17, HOIL-1 KO n = 5) or 100 (middle panel; control n = 10, HOIL-1 KO n = 10) tachyzoites T. gondii strain Pru-luc. (D) Log10 total flux (luciferase activity; photons per second) as a measure of parasite burden 8 days post-infection with 100 tachyzoites. Each symbol represents an individual mouse and the mean log10 is indicated. (E,F) Survival (E) and weight (F) of control (blue circles) and HOIL-1 KO (red squares) mice following oral gavage with 2 × 109 CFU C. rodentium. n = 20/group for survival and n = 10/group for weight. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001. Statistical analyses were performed using logrank Mantel–Cox test (A, C and E), Mann–Whitney test (B), or t-test (D).

DOI: http://dx.doi.org/10.7554/eLife.04494.006

Figure 1.

Figure 1—figure supplement 1. Comparison of RBCK1/HOIL1 alleles from RBCK1/HOIL1-mutant patients.

Figure 1—figure supplement 1.

Npl4 zinc finger ubiquitin binding domain, RING; really interesting new gene E3 ligase domain, IBR; Inbetween-RING domain. HOIL1L/RBCK1 isoform 2 (reference sequences NM_031229.2/NP_112506.2) was used for annotation.
DOI: http://dx.doi.org/10.7554/eLife.04494.007
Figure 1—figure supplement 2. Myocardium from aged HOIL-1 KO mice contains amylopectin-like deposits.

Figure 1—figure supplement 2.

Representative PAS (top), PAS plus diastase digestion (middle) and H&E-stained sections of myocardium from 18 month-old HOIL-1 KO (right) and control (left) mice. The scale bar (inset) represents 100 µm.
DOI: http://dx.doi.org/10.7554/eLife.04494.008
Figure 1—figure supplement 3. Analysis of hematopoietic cell populations from naïve HOIL-1 KO mice.

Figure 1—figure supplement 3.

(A–C) Flow cytometric analysis of cell populations in the spleen (A), thymus (B) and peritoneum (C) of HOIL-1 KO (red squares) and control (blue circles) mice. DP; CD4, CD8 double-positive. (D) Complete blood counts from HOIL-1 KO (red squares) and control (blue circles) mice. WBC, white blood cells, ×103/mm3, RBC, red blood cells, ×106/mm3; HGB, hemoglobin, g/dl; HCT, hematocrit, %; MCV, mean corpuscular volume, µm3; MCH, mean corpuscular hemoglobin, pg; MCHC, mean corpuscular hemoglobin concentration, %; Seg Neu, segmented neutrophils, %; Lymphos, lymphocytes, %, Monos, monocytes, %. Eosinophils, basophils or band neutrophils were not detected. Each symbol represents an individual mouse and the mean is indicated.
DOI: http://dx.doi.org/10.7554/eLife.04494.009
Figure 1—figure supplement 4. Hoil1/Rbck1 and neighboring gene (Trib3 and Tbc1d20) transcript expression in control and HOIL-1 KO bone marrow derived macrophages.

Figure 1—figure supplement 4.

Data represent the mean ± SEM from cells derived from four pairs of mice each analyzed in triplicate. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001. Statistical analyses were performed using t-test.
DOI: http://dx.doi.org/10.7554/eLife.04494.010
Figure 1—figure supplement 5. Pathology of HOIL-1 KO mice during Listeria infection.

Figure 1—figure supplement 5.

Alanine aminotransferase (ATL; left panel) and aspartate aminotransferase (AST; right panel) levels in serum from control (blue circles) and HOIL-1 KO (red squares) mice 6 days after infection with 105 CFU Listeria. Each symbol represents an individual mouse and the mean is indicated. *p ≤ 0.05, **p ≤ 0.01; t-test with Welch's correction.
DOI: http://dx.doi.org/10.7554/eLife.04494.011