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8 Figure S1. Generation of HspA4 -/- mice. (A) Structures of the wild-type (Wt) HspA4 -/- allele, targeting vector and targeted allele are shown together with the relevant restriction sites. The filled rectangles indicate the exons. Neo, DTA, and arrows (a and b) indicate pgk-neo selection cassette, MC1-DTA cassette, and the locations of primers for genomic PCR, respectively. X: XhoI, B: BamHI. (B) PCR genotyping of tail DNA of heterozygous (+/-) and homozygous (-/-) HspA4 knockout mice and wild-type (+/+) mice using primers specific to wild-type or mutant locus. (C) Expression of HspA4 protein. Figure S2. Generation of mouse embryonic fibroblast cell lines of HspA4 -/- genotype. (A) Expression of HspA4 protein in Apg-2-KO/Flp-In cells transfected with indicated vectors. (B) Expression of HspA4 protein in Apg-2-KO/Flp-In cells (HspA4 -/- ) with or without HSPA4 overexpression and wild type mouse embryonic fibloblasts (WT). Figure S3. (A) Association between HSPA4 and Bmi1 expression in the colonic mucosa of patients with Crohn s ileitis. Scatter plot of relative mrna levels of HSPA4 and the indicated gene in human colonic mucosa. (B) Relative expression levels of the indicated genes in the colonic mucosa of controls without IBD (control, colon) and the ileal mucosa of controls without IBD (control, small intestine), Crohn s ileitis (CD) and ulcerative colitis (UC). Figure S4. Receiver operator characteristics (ROC) analysis of the relative expression level of HSPA4 (A) and Bmi1 (B). The ROC analysis identified values of 0.98 and 2.2 as optimum cut-points to differentiate between patients in the high and low HSPA4 and

9 Bmi1 expression groups, respectively, for the prediction of response to steroid therapy. Figure S5. Representative immunohistochemical images of HSPA4 in human colitis-associated colorectal cancers. Scale bar, 100 µm. Figure S6. Relative expression of HSPA4 in different cell types. Lamina propria myeloid cells were sorted using immunomagnetic beads coated with monoclonal antibodies against CD4 and CD11b to isolate T cells and macrophages, respectively. The expression of HSPA4 mrna in colonic mucosa of WT mice untreated (control) or treated with DSS for 7 days was analyzed by quantitative real-time PCR. Figure S7. Enhanced apoptosis by HSPA4 knock-down in Jurkat cells. HSPA4 sirna or control sirna was transfected into Jurkat cells. After 12 hrs, cells were cultured with media containing thapsigargin (4 µm) or adriamycin (2 µm). (A, B) At 9 hrs after exposure to the stressors, Caspase-8 (A) and caspase-9 (B) activity was measured. (C) The relative number of viable cells was estimated by MTT assay at 17 hrs after exposure to the stressors. Data are expressed as the means + SEM. *P < 0.05 compared with non-treated cells

10 Supplementary Table 1 Primers for PCR RT-PCR was performed with the following sets of primers: for human β-actin, 5 -tccctggagaagagctacga-3 and 5 -aggaaggaaggctggaagag-3 ; for human HSPA4, 5 -caagcccaatggaaacatct-3 and 5 -ggatcagctccatcttctgc-3 ; for human Bcl-2, 5 - atgtgtgtggagagcgtcaa-3 and 5 -gccggttcaggtactcagtc-3 ; for human Bcl-xL, 5 -catggcagcagtaaagcaag-3 and 5 -tgggatgtcaggtcactgaa-3 ; for human Sox2, 5 -cgagtggaaacttttgtcgga-3 and 5 -tgtgcagcgctcgcag-3 ; for human Lgr5, 5 -gtcccacttcctgcatgtct-3 and 5 -ctcaggctcaccagatcctc-3 ; for human Bmi1, 5 -ccagggcttttcaaaaatga-3 and 5 -attagagccattggcagcat-3 ; for human Gankyrin, 5 -gggctgctggtgtcccaag-3 and 5 -attaaacccaggccaccttt-3 ; for human Cirp, 5 -gagggctgagttttgacacc-3 and 5 -tgggtctccctgtctttcac-3 ; for mouse β-actin, 5 -agcgaagagctacaggcaag-3 and 5 -tcacacacaggcaccttctc-3 ; for mouse Bcl-2, 5 -aagctgtcacagaggggcta-3 and 5 -caggctggaaggagaagatg-3 ; for mouse Bcl-xL, 5 -ttcgggatggagtaaactgg-3 and 5 -tgcaatccgactcaccaata-3 ; for mouse Sox2, 5 -gaacgccttcatggtatggt-3 and 5 -ttgctgatctccgagttgtg-3 ; for mouse Lgr5, 5 -aacggtcctgtgagtcaacc-3 and 5 -agtcatggggtaagctggtc-3 ; and for mouse Cirp, 5 -gaggactcagcttcgacacc-3 and 5 -ctccctgtcctttaccacca-3.

11 Supplementary Table 2. Comaprison between steroid-sensitive and resistant patients with ulcerative colitis Steroid-sensitive Steroid-resistant P-value patients (n=15) patients (n=8) Gender (male/female) 5/10 2/6 1.0 Age (year) Disease duration (year) C-reactive protein (mg/dl) Hemoglobin (g/dl) Matts score HSPA4 expression * Bmi1 expression * In addition to clinical parameters, relative HSPA4 and Bmi1 mrna expression in colonic mucosae from steroid responders and non-responders was compared. Difference in gender was analyzed by Fisher s exact test. *P < 0.05