The frequencies of haplotype of KRT1 gene in SLE, SSc and control group.

<p>The frequencies of haplotype of KRT1 gene in SLE, SSc and control group.</p

The genotyping results of samples by PCR-SSP.

<p>(a): Typing for SNP rs14024 is based on the two PCR reactions using specific sequence primers, SNP with A or G, or both were designed with a size band of 410 bp in PCR; a 689 bp band serves as the internal positive control. (b): The 21bp nucleotide deletion was detected with a small band (S) and wild type of rs267607656 is demonstrated with a large band (L) in PRC products. The PCR results of three control samples (con1, con2 and con3) and object samples (S1, S2, S3 et al.) are given, following by the read of typing results in parenthesis.</p

The allele frequencies of SNP rs14024 and indel rs267607656 in different populations.

<p>The allele frequencies of SNP rs14024 and indel rs267607656 in different populations.</p

The genotype frequencies of SNP rs14024 and indel rs267607656 in different populations.

<p>The genotype frequencies of SNP rs14024 and indel rs267607656 in different populations.</p

Validation of PCR-SSP genotyping assay.

<p>6 reference DNA samples identified with Sanger sequence-based typing (SBT) are utilized to validate the new method of PCR-SSP typing. (a): PCR products with KRT1 exon 9 DNA sequences were sequenced with forward and reverse sequencing primers. The 21-bp deletion and non-deletion in 6 samples are labeled in the left profiles of reverse-primer sequencing. The SNPs rs14024 with A, G or both (A/G) at position 388 in exon 9 of KRT1 gene are given in the right side. The same samples were used to type by PCR-SSP. (b): A 689 bp band, amplified by GAPDH primers serves as a positive control. Typing of SNP with either A, G or both at rs 14024 is read based on the presence of a 410-bp band in PCR products in different primer pairs. (c): The indel polymorphism in rs267607656 of KRT1 is detected by the sizes of PCR gels with larger bands (L, no deletion) and/or small bands (S, with deletion). The relatively small size (S) detected is stand for the existing of a 21-bp deletion in KRT1 gene.</p

Antibodies against HLA/MICA antigens in pre- and post-transplant serum samples.

<p>IgG antibodies against HLA class I and II antigens and MICA antigens were measured by single antigen Luminex beads arrays. MFI values represent the quantitation of alloantibodies detected. (A) The donor’s and recipient’s HLA antigen types and the positive reacted HLA antigens (others) with test serum samples are labeled as grouped.The MFI values of antibodies detected from patient serum before (gray bars) and after the first transplant (solid bar) are given. (B) Anti-MICA antibodies were detected using 11 common MICA antigens. MICA001, MICA002, MICA007, MICA012, MICA017, and MICA018, all from the MICA-G1 antigen group were present in pre- and post-first transplant patient sera; other antigens tested were negative. DSA to the antigen produced by MICA*018 allele is indicated with an arrow. C1q-PE, rather than goat anti human IgG conjugated with PE, was used to demonstrate the complement fix (C1q, white bars).</p

C4d deposition in the walls of glomerular capillaries of allograft.

<p>Immunofluorescent microscopic magnification×400.</p

MICA expressed on HUVEC cell surfaces is the target for anti-MICA antibodies.

<p>(A) HUVECs were freshly isolated from five umbilical cord samples. Cells were stained with mAb W6/32 for HLA class I antigens (solid profile), 6B3 for MICA antigens (open profile; dark line) and normal mouse IgG as control (open profile; light line). (B). HUVECs were incubated with normal human serum (NHS), pooled PRA+ sera (POS), or patient serum (after first transplant, with platelet absorption). Cytotoxicity is reported as percent lysis.</p

HLA/MICA typing results.

<p>HLA/MICA typing results.</p