EMBO/soda-nel huggingface dataset:
Inhibition of lethal inflammatory responses through the targeting of membrane-associated Toll-like receptor 4 signaling complexes with a Smad6-derived peptide. (2015)
Figure 4

Smaducin-6 is therapeutic for mice with CLP-induced sepsis

Subcutaneous injection of Smaducin-6 increases the survival rate of severe CLP-induced sepsis mice (BALB/c mice). Different amounts of Smaducin-6 were subcutaneously injected at 2 h post-CLP followed by three injections at 12-h intervals. n = 10 mice per group per experiment.

Smaducin-6 treatment reduces abnormal nuclei and cell morphology observed in tissues of CLP-induced BALB/c mice, as observed by hematoxylin and eosin (H/E) staining. Data shown are representative of five independent experiments. Scale bar, 40 μm (magnification, 400×).

The survival rates of severe CLP-induced sepsis mice by subcutaneously injected Smaducin-6 were compared with those by intraperitoneally injected antibiotics (8 mg/kg gentamycin plus 8 mg/kg cephalosporin). Antibiotics were intraperitoneally injected into CLP mice at 2 h post-CLP followed by an additional injection after 12 h. n = 10 mice per group per experiment.

Therapeutic effects of Smaducin-6 alone, or antibiotics (8 mg/kg gentamycin plus 8 mg/kg cephalosporin), or different amounts of Smaducin-6 plus antibiotics were observed in CLP mice. Antibiotics were intraperitoneally injected into CLP mice at 2 h post-CLP followed by an additional injection after 12 h. Smaducin-6 was subcutaneously injected at 2 h post-CLP followed by three injections at 12-h intervals. n = 10 mice per group.

Comparison of mortality of severe CLP-induced sepsis mice upon subcutaneous injections of a total of 16 mg/kg Smaducin-6 or TAT-S6(422-441) peptide. n = 10 mice per group.

Data information: In (A–E), PBS and scrambled peptide (Pal-Scram #1) were subcutaneously injected into CLP mice, respectively, as negative controls. Data in (A–E) were statistically analyzed by the log-rank test. ***P < 0.001, **P < 0.005, *P < 0.05 compared to vehicle control (CLP + Pal-Scram #1 or CLP + PBS).
Inhibition of lethal inflammatory responses through the targeting of membrane-associated Toll-like receptor 4 signaling complexes with a Smad6-derived peptide. (2015)
Figure 5

Smaducin-6 reduces systemic pro-inflammatory cytokines in CLP-induced sepsis mice

A–I Smaducin-6 treatment decreased the concentration of IL-6, TNF-α, IFN-γ, and IL-1β in the blood of CLP mice (A–D), but not IL-4, IL-10, TGF-β1, IL-12, or IL-17A (E–I). Cytokine concentrations were analyzed by ELISA. n = 10 mice per group per experiment. Data were statistically analyzed by the Dunnett's multiple comparison test (one-way ANOVA). ***P < 0.001, **P < 0.005, *P < 0.05 compared to sham or vehicle control (CLP + Pal-Scram #1).

J Expression of the IL-6 protein is reduced in the spleen and liver of Smaducin-6-treated CLP mice. Each lane represents independent mice.

Source data are available online for this figure.
Inhibition of lethal inflammatory responses through the targeting of membrane-associated Toll-like receptor 4 signaling complexes with a Smad6-derived peptide. (2015)
Figure 6

Smaducin-6 shows bactericidal effects through neutrophil recruitment

Subcutaneous injection of Smaducin-6 into CLP mice reduced bacterial loads in peritoneal fluid. n = 10 mice per group per experiment. Data were statistically analyzed by the Mann–Whitney U-test. **P < 0.005 compared to sham or vehicle control (CLP + Pal-Scram #1).

Smaducin-6 increases neutrophil numbers in peritoneal lavage fluids from LPS-induced endotoxemia BALB/c mice. n = 5 mice per group per experiment. Data were statistically analyzed by a t-test. **P < 0.005 compared to sham or vehicle control (CLP + Pal-Scram #1).

CXCR2 expression in human neutrophils, measured by FACS, was decreased by LPS treatment and restored by Smaducin-6 treatment.

Treatment with Smaducin-6 downregulates LPS-induced GRK2 expression in RAW264.7 cells and human neutrophils.

CXCR2-expressing neutrophils isolated from CLP mice in the presence of Smaducin-6 or Pal-Scram peptide were analyzed by FACS. n = 5 mice per group were used. Data were statistically analyzed by a t-test and show the mean ± SD of at least three independent experiments. **P < 0.005 compared to sham or vehicle controls (CLP + PBS and CLP + Pal-Scram #1).

GRK2 expression was directly analyzed by immunoblotting in neutrophils isolated from CLP mice in the presence of Smaducin-6 or Pal-Scram #1 peptide.

Data information: Data in (C), (D), and (F) are representative of three independent experiments. Source data are available online for this figure.
Inhibition of lethal inflammatory responses through the targeting of membrane-associated Toll-like receptor 4 signaling complexes with a Smad6-derived peptide. (2015)
Figure 7

Smaducin-6 reduces sepsis-induced apoptosis

A, B Quantitative analysis of (A) TUNEL-positive cells and (B) caspase-3-positive cells in IHC in the spleens of CLP + PBS, CLP + Pal-Scram #1, and CLP + Smaducin-6 mice. Three independent experiments (n = 3 mice per group per experiments) were performed. At least five hot spots in a section of TUNEL and IHC per experiment were selected, and average count was determined. The data were expressed as a mean percentage of total cell numbers and statistically analyzed by a t-test and show the mean ± SD of three independent experiments. **P < 0.005, ***P < 0.001 compared to sham or vehicle control (CLP + Pal-Scram #1).

C ELISA showing that Smaducin-6 treatment reduced IFN-β1 levels in peritoneal lavage fluids compared to Pal-Scram #1-treated CLP mice. n = 10 mice per group per experiment. Data were statistically analyzed by the Dunnett's multiple comparison test (one-way ANOVA). ***P < 0.001 compared to sham or vehicle control (CLP + Pal-Scram #1).

D, E IFN-β1 (D) and TRAIL (E) expression detected by FACS in splenocytes is decreased in Smaducin-6-treated CLP mice compared to scrambled peptide-treated CLP mice. Data are representative of three independent experiments.

F Decreased expression of TRAIL was detected by IHC in the spleen of CLP-induced mice treated with Smaducin-6 compared to mice treated with Pal-Scram. Scale bar, 100 μm (magnification, 200×). Data are representative of three independent experiments.
Nhp2 is a reader of H2AQ105me and part of a network integrating metabolism with rRNA synthesis (2021)
Figure 1

Figure 1: H2AQ105me fluctuates with the cell cycle and is a mark of proliferating cells. A) Cell cycle profile of H2AQ105me. Logarithmically growing cells were arrested using alpha-factor for 2.5 hours. Upon release (t=0), samples were taken every 15 minutes and samples were analysed for H2AQ105me levels using western blotting. S-phase is indicated by the enrichment of H3K56ac.

B) H2AQ105me levels upon arrest (2.5 hours) at different cell cycle stages. Treatment is given above the lanes.

C) H2AQ105me levels upon entry of cells into stationary phase.

D) H2AQ105me levels in various histone mutations targeting sites of well-known cell cycle-dependent modifications.

E) H2AQ105me and H3K56ac levels in deletions of genes important for H3K56ac deposition (see text for details).

F) Scheme of rDNA locus and indication of primer locations used in G-J.

G-J) ChIP-qPCR for indicated antibodies after treatment with 5nM rapamycin for the indicated time points. Samples were either normalized to input or to the core histone. n=3, biological replicates, error bars are standard error of the mean. *, p<0.05, relative to t=0, based on unpaired, 2-tailed t-test

K) ChIP-qPCR for Nop1-FLAG using anti-FLAG tag antibody. Samples were normalized using input. n=3, biological replicates, error bars are standard error of the mean. **, p<0.001, based on unpaired, 2-tailed t-test
Nhp2 is a reader of H2AQ105me and part of a network integrating metabolism with rRNA synthesis (2021)
Figure 2

Figure 2: Mass spectrometry identifies ribosome biogenesis factors to be recruited to H2AQ105me.

A) Schematic of the experimental strategy.

B) Volcano plot of quantitative SILAC experiments. Left upper quadrant indicates significantly, 2-fold enriched proteins bound specifically to H2AQ105me. Members of the FACT complex (SPT16/POB3) are highlighted in red.

C) STRING network of proteins enriched on H2AQ105me (highlighted in red are proteins involved in ribosome biogenesis). Protein names can be found in Dataset EV1.

D) Gene ontology enrichment for proteins bound to H2AQ105me.

E) Gene set enrichment analysis for the altered transcriptome in an H2AQ105A background compared to wildtype. The normalized enrichment score (NES) is reported as are individual p-values. Vertical black lines on the x-axis represent genes in gene sets. The green line connects points of enrichment score and genes. Enrichment score is the maximum deviation from zero as calculated for each gene going down the ranked list, and represents the degree of over-representation of a gene set at the top or the bottom of the ranked gene list.
Nhp2 is a reader of H2AQ105me and part of a network integrating metabolism with rRNA synthesis (2021)
Figure 3

Figure 3: Nhp2 is an epigenetic reader of H2AQ105me.

A) ChIP-qPCR for Nhp2-myc using anti-myc antibody across the rDNA locus. Samples were normalized to input. n=3, biological replicates, error bars are standard error of the mean. *, p<0.05, **, p<0.001, based on unpaired, 2-tailed t-test

B) Peptide pulldown on H2AQ105 peptides from yeast expressing myc-tagged Nhp2.

C) Peptide pulldown using recombinantly purified Nhp2. Eluted peptides were slot blotted and detected using streptavidine-HRP.

D) Peptide pulldown on H2AQ105 peptides from lysates of human cells (HEK293T).

E) ChIP-seq profile of Nhp2 (orange) and its respective input (black) across the entire chromosome XII of yeast. H2A-normalised H2AQ105me (grey) was directly taken from (Tessarz et al., 2014) as comparison. Magnification shows the two annotated rDNA loci in the yeast genome indicated by the black bars below the tracks. Scale bar is for the magnification.
Nhp2 is a reader of H2AQ105me and part of a network integrating metabolism with rRNA synthesis (2021)
Figure 4

Figure 4: Nhp2 bridges components of the SSU and H2AQ105me.

A) Peptide pulldowns from indicated TAP-tagged strains. Pulldown efficiency was analyzed by western blotting.

B) Spot test of the indicated strains on YPD or YPD supplemented with the given antibiotics. Overnight yeast cultures were adjusted to OD600=1 and then serially diluted 5-fold before spotting on indicated media.

C) Ribosome slippage assay using a tandem luciferase reporter separated by a sequence allowing for slippage. The sequences allowing for slippage are given below the graph. n=8, biological replicates, error bars are standard error of the mean. Significance was tested using 2-tailed, unpaired t-test.

D) Proteomic analysis of Nhp2 interactors. Highlighted in red are the known members of the H/ACA pseudouridylation complex. The red dashed line indicates significance cutoff (FDR < 0.05; for a full list of enriched proteins, please refer to Dataset EV3).

E) Intersection of Nhp2 and H2AQ105me pulldown. 17 proteins were enriched in both experiments and the majority are members of the small subunit processome (highlighted in red).

F) GO enrichment for these 17 commonly identified proteins shown in Figure 4E.

G) Peptide pulldowns from human cells (HEK293T) to test for conservation of the reported interactions between Nhp2 and members of the SSU processome complex on H2AQ105me. Dyskering (Cbf5 homolog) was probed to test for the presence of members of the H/ACA complex. Pulldown efficiency was analyzed by western blotting using the indicated antibodies.

H) Simplified schematic overview over some processing steps of the rRNA. Cut sites are indicated as A0-A3. Blue bars indicate Northern probes to detect 18S and 25S mature rRNA, respectively (please also see Figure EV4). Labelled bars on top of the scheme indicate used probes. For a representative Northern Blot, please be referred to the Source Data accompanying Figure 4.

I) Quantification of Northern Blot results. Normalised to GAPDH, n=3, error bars are standard error of the mean. Asterix indicates p-value (*, p<0.05, based on unpaired, 2-tailed t-test).

EXPANDED VIEW LEGENDS
Spatial control of lipid droplet proteins by the ERAD ubiquitin ligase Doa10 (2016)
Figure 1

Figure 1. The LD protein Pgc1 is an ERAD substrate.

(A) The degradation of 3HA-Pgc1 was analyzed in cells with the indicated genotype upon inhibition of protein synthesis with cycloheximide (chx). A plasmid-borne 3HA-Pgc1 expressed from the endogenous promoter was used. 3HA-Pgc1 was detected with anti-HA antibodies. Kar2 was used as a loading control and detected with anti-Kar2 antibodies. The graph on the right shows the average of four independent experiments; error bars represent the standard deviation.

(B) The ubiquitination of 3HA-Pgc1 was analyzed in cells with the indicated genotype and expressing myc-tagged ubiquitin. 3HA-Pgc1 was immunoprecipitated using anti-HA and polyubiquitin-conjugated 3HA-Pgc1 was detected using anti-myc antibody. Input fraction corresponds to 2% of the total amount used for IP.

(C) The degradation of 3HA-Pgc1 was analyzed as in (A) in cells bearing the CDC48 temperature-sensitive allele cdc48-6. Cells were grown 2,5 hours at 37C prior addition of cycloheximide. Inactivation of Cdc48 mutant protein was confirmed by stabilization of the ERAD substrate Erg1 in the same cells.

(D) Membrane association of ubiquitinated 3HA-Pgc1 was analyzed by density gradient centrifugation followed by immunoprecipitation. Cells of the indicated genotype and expressing myc-tagged ubiquitin were grown as in (C). Lysates were subjected to centrifugation on an optiprep density gradient. Collected fractions were analyzed by western blotting before and after immunoprecipitation with anti-HA antibodies. Dpm1 and Pgk1 were used as markers for membrane bound and soluble fractions respectively. Input fraction corresponds to 10% of the total amount used for IP. “Top”, “mid”, “bottom” indicate the fractions from the gradient. Asterisk marks a faster migrating unspecific band in the input bottom fraction.
Spatial control of lipid droplet proteins by the ERAD ubiquitin ligase Doa10 (2016)
Figure 2

Figure 2. Pgc1 behaves as an integral membrane protein.

(A) Localization of chromosomally expressed GFP-Pgc1 from the constitutive ADH1 promoter in wt and doa10Δ cells. Arrowheads indicate GFP-Pgc1 labeling at the ER, which is stained by Sec63-Cherry. LDs were visualized upon staining with the neutral lipid dye MDH. On the bottom, box plot with quantitation of GFP-Pgc1 fluorescence intensity at the nuclear envelope in wt and doa10Δ. GFP-Pgc1 measurements were performed as described in the Materials and Methods section. Scale bar: 5 µm.

(B) Schematic representation of Pgc1. The location of the predicted hydrophobic domain is indicated.

(C) Crude membranes from wt and doa10Δ cells expressing 3HA-Pgc1 were subjected to the indicated treatments and subsequently fractionated into membrane pellet (P) and supernatant (S).

(D) Crude membranes from wt and doa10Δ cells expressing 3HA-GFP-Pgc1275-321 were analyzed as in (C).
Spatial control of lipid droplet proteins by the ERAD ubiquitin ligase Doa10 (2016)
Figure 3

Figure 3. Pgc1 localizes to the ER before stably concentrating on LDs.

(A) Localization of GFP-Pgc1 in are1Δ are2Δ lro1Δ dga1Δ cells in presence (no LDs) or absence of DOA10 (no LDs doa10Δ). The ER was visualized by expression of Sec63-Cherry. Scale bar: 5 µm.

(B) GFP-Pgc1 was expressed under the ADH1 promoter; LD formation was stimulated by galactose-induced expression of DGA1 in are1Δare2Δlro1Δ. Fluorescence microscopy was used to follow GFP-Pgc1 localization over time. LDs were visualized upon staining with the neutral lipid dye MDH. Scale bar: 5 µm.

(C) Photo-conversion of tdEOS-Pgc1 expressed under the ADH1 promoter; LD formation was stimulated by galactose-induced expression of DGA1 in are1Δare2Δlro1Δdoa10Δ cells. The red square marks the photo-converted region. The time (in minutes) after photoconversion is indicated. Arrowheads point LDs containing photoconverted tdEOS-Pgc1. Scale bar 5µm.

(D) FRAP experiment of GFP-Pgc1 in wt and doa10Δcells. Representative examples are shown. The bleached areas are marked by red squares and include a LD adjacent to ER. The time (in seconds) after photobleaching is indicated. Each graph shows average fluorescence intensities for 10 cells normalized to pre-bleached plotted over time. Error bars indicate standard deviation.
Spatial control of lipid droplet proteins by the ERAD ubiquitin ligase Doa10 (2016)
Figure 4

Figure 4. Pgc1 is degraded by Doa10 at the ER.

(A) The degradation of 3HA-Pgc1 was analyzed in cells with the indicated genotype as in Fig. 1A. The graph shows the average of three independent experiments; error bars represent the standard deviation.

(B) The degradation of the Doa10 substrates 3HA-Pgc1 and 3HA-Vma12-Ndc10C’ was analyzed in cells of the indicated genotypes treated with oleic acid. 3HA-Pgc1 and 3HA-Vma12-Ndc10C’ were detected with anti-HA antibodies. Dpm1 was used as a loading control and detected with anti-Dpm1 antibodies. The graph shows the average of three independent experiments; error bars represent the standard deviation.
Spatial control of lipid droplet proteins by the ERAD ubiquitin ligase Doa10 (2016)
Figure 5

Figure 5. Pgc1 hydrophobic hairpin is necessary and sufficient for Doa10-dependent degradation.

(A) Localization of GFP-tagged Pgc1 derivatives in which the hydrophobic hairpin (aa275-321) was replaced by membrane anchor of Scs2 (Pgc1Scs2MA) or Bos1 (Pgc1Bos1MA). ER and LDs were visualized by expression of Sec63-Cherry and MDH staining, respectively. Scale bar: 5 µm.

(B) The degradation of 3HA-Pgc1 or the indicated chimeras in wt and doa10Δ cells was analyzed as in Fig. 1A. The blot is representative of at least three independent experiments.

(C) The degradation of 3HA-GFP-Pgc1275-321 in wt and doa10Δ cells was analyzed as in Fig. 1A. The graph shows the average of four independent experiments; error bars represent the standard deviation.

(D) Localization of 3HA-GFP-Pgc1275-321 in doa10Δ cells. The ER was visualized by expression of Sec63-Cherry. Scale bar: 5 µm.
Spatial control of lipid droplet proteins by the ERAD ubiquitin ligase Doa10 (2016)
Figure 6

Figure 6. A heterologous hydrophobic hairpin functions as a Doa10 degron.

(A) The degradation of 3HA-Pgc1-GPAT160-216, containing GPAT4 hydrophobic hairpin (aa160-216) replacing the one of Pgc1 was analyzed in cells with the indicated genotype as in Fig. 1A. The graph shows the average of three independent experiments; error bars represent the standard deviation.

(B) Localization of GFP-Pgc1-GPAT4160-216 in cells with the indicated genotype. Arrowheads indicate GFP-Pgc1-GPAT4160-216 labeling at the ER, marked with Sec63-Cherry. LDs were visualized upon staining with MDH. Scale bar: 5 µm.

(C) The degradation of 3HA-Pgc1-GPAT160-216 in cells with the indicated genotype was analyzed as in Fig. 1A. The graph shows the average of three independent experiments; error bars represent the standard deviation.

(D) Localization of GFP-Pgc1-GPAT160-216 in are1Δ are2Δ lro1Δ dga1Δ doa10Δ mutant (no LDs doa10Δ). The ER was visualized by expression of Sec63-Cherry. Scale bar: 5 µm.
Zinc inhibits lethal shock by reducing microbe-induced interferon signature in intestinal epithelium (2020)
Figure 1

Figure 1: ZnSO4 fails to protect against a lethal TNF injection in Adx, GRDim and GRVillKO mice.

(A) Survival curves of C57BL/6J naïve and adrenalectomized (Adx) mice treated for 7 days with 25 mM ZnSO4 in the drinking water and challenged, i.p., with 30 µg (naïve) or 2 µg (Adx) TNF, solved in sterile PBS, per 20 g body weight. No deaths occurred later than 72 h after TNF injection. Results represent combined data of 3 experiments.

(B) Corticosterone levels in the serum of C57BL/6J naïve and Adx mice after 7 days of 25 mM ZnSO4 supplementation to the drinking water (N=6 per group).

(C) Survival curves of GRWT and GRDim mice treated for 7 days with 25 mM ZnSO4 in the drinking water and challenged, i.p., with 50 µg (GRWT) or 12.5 µg (GRDim) TNF, solved in sterile PBS, per 20 g bodyweight. No deaths occurred later than 50 h after TNF injection. Results represent combined data of 2 experiments.

(D-E) Zinc levels in the serum after 7 days of 25 mM ZnSO4 supplementation to the drinking water of C57BL/6J naïve and Adx mice (D, N=3-6), or GRWT and GRDim mice (E, N=6-9).

(F) Agtpbp1 gene-expression measured with RT-qPCR in the ileum of GRWT and GRDim mice treated for 7 days with 25 mM ZnSO4 in the drinking water (N=3 per group).

(G) Survival curves of GRfl/fl and GRVillKO mice treated for 7 days with 25 mM ZnSO4 in the drinking water and challenged, i.p., with 35 µg TNF, solved in sterile PBS, per 20 g bodyweight. No deaths occurred later than 150 h after TNF injection. Results represent combined data of 6 experiments.

(H) Mouse Nr3c1 gene-expression measured with RT-qPCR in the intestinal epithelial cells (IECs) of GRfl/fl and GRVillKO mice (N=3-6). Hprt and Villin were used as house-keeping genes.

(I) Zinc levels in the serum after 7 days of 25 mM ZnSO4 supplementation to the drinking water of GRfl/fl and GRVillKO mice (N=9 per group).

(J) Number of genes differentially expressed (up and down) by zinc (LFC>0.8 and p<0.05) measured by RNA sequencing in the ileum and liver of C57BL/6J mice treated with 25 mM ZnSO4 in the drinking water for 7 days (N=3).

Data information: For the survival curves p-values were analyzed with a Chi2-test. In (B,D,E,F,H,I) the data is shown as mean +/- SD. p-values were analyzed with a student's t-test (unpaired, two-tailed) on the log transformed data in (B,H) and with a two-way ANOVA followed by a Tukey multiple comparisons test in (D,E,F,I). Significant expression of genes in the RNA sequencing was assessed with a Wald test with negative binomial distribution in DESEQ2.
Zinc inhibits lethal shock by reducing microbe-induced interferon signature in intestinal epithelium (2020)
Figure 2

Figure 2: Transcriptional effects of ZnSO4 in the ileum and the role antibiotics and germ free aspects.

(A-C) RNA sequencing on ileum samples of C57BL/6J mice treated with water or 25 mM ZnSO4 water for 7 days and then injected i.p. with 200 µg Dex (Rapidexon) solved in 200 µL PBS, or with PBS only. 2h after injection the ileum was isolated (N=3 per group). Overlap between genes (A) upregulated or (B) downregulated by ZnSO4 (Zn) or Dex (LFC>0.8, p<0.05). (C) Scheme of numbers of genes upregulated in ZnSO4, Dex or ZnSO4+Dex treated mice (LFC>0.8, p<0.05). Significant expression of genes in the RNA sequencing was assessed with a Wald test with negative binomial distribution in DESEQ2.

(D) Regression curve plotting LFCs of genes induced by ZnSO4 in GRWT and GRDim mice, as measured by RNA-seq in ileum samples. All genes, induced in ileum by DEX in GRWT with LFC>0.8, p<0.05 are considered, while these genes with p<0.05 in GRDim were considered. The slope of the correlation curve as well as Pearson correlation coefficient were calculated by Graphpad Prism.

(E) C57BL/6J mice received antibiotics (AB) in the drinking water for 3 weeks, followed by 1 week 25 mM ZnSO4 in the drinking water. During this week AB administration was continued by oral gavage. Mice were challenged i.v. with 12.5 µg TNF solved in sterile PBS and survival recorded. Combined data of 2 experiments.

(F) Zinc levels measured in serum after the antibiotics and ZnSO4 protocol (N=5-8).

(G-H) Mice housed in germ-free (GF) or specific pathogen free (SPF) conditions received 25 mM ZnSO4 in the drinking water for 1 week and were then challenged i.p. with 35 µg TNF, dissolved in sterile PBS, per 20 g bodyweight. Combined data of 2 experiments. (H) displays the blood zinc levels in SPF and GF mice treated for 1 week with water or ZnSO4 (N=3/4 per group).

(I) Ileum Slurry (IS) was isolated from C57BL/6J mice put on 25 mM ZnSO4 or control water for 1 week and 750 µL of this IS was injected i.p. in C57BL/6J GF mice.

Data information: For the survival curves p-values were analyzed with a Chi2-test. In F and H data are shown as mean +/- SD and p-values were analyzed with a two-way ANOVA.