PMC

To calculate lab Jaccard distances between two labs, such as lab A and lab C, we first build a fragment set, F_A and F_C, for each lab. A fragment set contains all the pan-genome fragments annotated by the corresponding labs. The lab Jaccard distance between lab A and lab C is JD(A, C) = 1 − J(F_A, F_C), where the Jaccard index (J(F_A, F_C)) is the fraction of shared pan-genome fragments out of all the different fragments lab A and C have. A large lab Jaccard distance indicates two labs have few shared sequences.

(A) Cytokine production. LAB^+/+, LAB^−/−, and LAB^m/m BMDCs were purified and seeded in a 96-well plate and were stimulated with different concentrations of LPS for 16 hrs before ELISA. Each value is the mean and standard deviation of data from triplicate samples. Data with significant differences (p<0.05 for two-tailed t-test) are indicated by *. Data shown are representative of three experiments.(B) The absence of LAB phosphorylation. LAB^+/+, LAB^−/−, and LAB^m/m BMDCs were stimulated with LPS for 10 mins or pervanadate for 2 mins before lysis. LAB was immunoprecipitated and blotted with anti-pTyr and anti-LAB antibodies. (C) Hyperphosphorylation of Syk in LAB^−/− DCs. BMDCs were stimulated with 100ng/ml LPS for 30 minutes before lysis in RIPA buffer. The lysates were analyzed by immunoblotting with anti-pTyr, phospho-Syk (pY352), and Syk antibodies. Data shown are representative of three experiments.(D) The effect of LAB deficiency and LAB 5YF mutation on LPS-mediated MAPK activation. BMDCs from LAB^−/−, LAB^+/+ and LAB^m/m mice were stimulated with LPS before lysis at the indicated time points. The lysates were analyzed by blotting with phospho-specific or pan-antibodies against ERK, p38, and JNK. Data shown are representative of three experiments. Numbers shown are relative intensities for phosphorylated ERK, p38, and JNK1 normalized to their pan forms.

Pearson product-moment correlation was calculated to examine differences in UA concentrations measured by the two different laboratories. (A) Correlation of ACDua from the same blood sample measured by Lab A and Lab B (R²=0.5337, P<0.01). Mean percent difference between Lab A ACDua and Lab B ACDua was 38.5%. (B) Correlation of SSTua from the same blood sample measured by Lab A and Lab B (R²=0.9584, P<0.01). Mean percent difference between Lab A SSTua and Lab B SSTua was 4.9%.

(A)Septic shock. After injection of LPS, the survival of LAB^m/m, LAB^−/− and LAB^+/+ mice(eight mice for each genotype) was monitored for up to 30 hrs. The survival of these mice after LPS treatment was analyzed statistically by the Kaplan-Meyer method (log-rank test: LAB^−/− vs. LAB^+/+, p<0.0001; LAB^−/− vs. LAB^m/m, p=0.04; LAB^m/m vs. LAB^+/+, p=0.07).(B) LPS-induced cytokine production in vivo. LAB^m/m, LAB^−/−, and LAB^+/+ mice (4 each) were injected as above. Sera were collected at the indicated time points and cytokine concentrations were determined by ELISA. Each value is the mean and standard deviation of data from four mice. Data with significant difference (p<0.05 and p<0.01 for two-tailed t-test) are indicated by * and **. Data shown are representative of three experiments.