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Figure S1. Enrichment of human CD22+ B cells. Peripheral blood mononuclear cells (PBMCs) were purified by density gradient centrifugation from peripheral blood. B cells were isolated with magnetic anti-CD22 beads (MACS). Purity of B cells was analyzed by staining with FITC-conjugated anti-CD19 antibody and FACS. Top right panel: depleted fraction, lower right panel: enriched fraction.

Figure S2. miR-24–3p knockdown in U266 and JK-6L cells. (A),(B) Viability after miR-24–3p knockdown. U266 (A) and JK-6L cells (B) were transfected with either anti-miR-24–3p-siRNA (miR-24–3p siRNA) (n = 16) or negative control-siRNA (control siRNA) (n = 3) for 24 h (U266, n = 6) and 48 h (JK-6L, n = 11). Viability (%) determined with trypan-blue staining normalized to medium control (med.). (C) control qPCR of miR-24–3p expression normalized to RNU1–1 from RPMI-8226 cells after transfection with either negative control siRNA (control siRNA) or miR-24–3p-specific siRNA (miR- 24–3p siRNA). Relative expression levels were calculated by the __ cT method with negative control as reference (set as 1). (D) qPCR analysis of bim expression normalized to beta-actin from U266 and JK-6L cells transfected with anti-miR-24–3psiRNA (miR-24–3p siRNA) (n = 9) or negative control-siRNA (control siRNA) (n = 9) for 24 or 48 h. (E), (F) qPCR analysis of potential apoptosis related targets of miR-24–3p faf1 (FAS-associated factor 1) and xiap (X-linked inhibitor of apoptosis protein) normalized to beta-actin from RPMI-8226, U266 and JK-6L cells transfected with anti-miR-24–3p-siRNA (miR-24–3p siRNA) (n = 9) or negative control-siRNA for 24 or 48 h. Relative expression levels were calculated by the __ cT method with negative control as reference (set as 1). (G) Proliferation rate (%) in RPMI-8226 cells labeled with 1nM CFDA-SE following transfection with anti-miR-24–3p-siRNA (miR-24–3p siRNA), negative control-siRNA (control siRNA) and FACS-analysis after 48 h. Reference: medium cultured RPMI-8226 cells (med.). Mean ± SEM. n = 3.

Figure S3. Proliferation level after stimulation with IL-6 or SDF-1α. RPMI-8226 cells labeled with 1nM CFDA-SE followed by stimulation with 10 ng/ml, 50/ml ng and 100 ng/ml rh-IL-6 (A) or SDF-1α (B) subsequent FACS-analysis after 24, 48, and 72h. Reference: naïve RPMI-8226 cells (black bars). Mean±SEM. n = 4. (C) Representative histograms. (D) Quantitative analysis of miR-24–3p expression in U266 and JK-6L normalized to RNU1–1. Cells were stimulated with 10 ng/ml, 50 ng/ml and 100 ng/ml rh-IL-6 for 24 h. Relative expression levels were calculated by the __ cT method with negative control as reference (set as 1). Mean values ± SEM; ** p<0.01; *** p<0.001.n = 9

Figure S4. IL-6 mediated survival of PC under ER-stress conditions. RPMI-8226 cells treated with 0.25 μg/ml tunicamycin (Tunica.) and 50 ng/ml rh-IL-6 (Tunica.+IL- 6), or 50 ng/ml rh-SDF-1α (Tunica.+SDF-1 α) for 24 h (A) and 48 h (B). Trypan-blue staining normalized to naïve cells (naive). n = 6. RPMI-8226 (C) and JK-6L cells (D) treated with 0.25 μg/ml tunicamycin (Tunica.) and 50 ng/ml rh-IL-6 (Tunica.+IL-6) followed by annexin-V/PI staining and FACS analysis. Naïve cells were used as control. Representative histograms. (E) Viability of JK-6L cells after treatment with 0.25 μg/ml tunicamycin (Tunica.) or 50 ng/ml rh-IL-6 (Tunica. + IL-6) for 48 h. Viability (%) was determined using trypan-blue staining normalized to naïve cells (naive). n = 12. qPCR analysis of miR-24–3p expression normalized to RNU1–1 (F) and bim expression normalized to beta-actin (G) from JK-6L cells treated with 0.25 μg/ml tunicamycin (Tunica.) alone or together with 50 ng/ml rh-IL-6 (Tunica. + IL-6) for 48 h. Relative expression levels were calculated by the __ cT method. n = 6. (H) Viability of JK-6L cells treated with 0.25 μg/ml tunicamycin alone (n = 6) or together with 50 ng/ml rh-IL-6 (n = 6) for 48 h following transfection with anti-miR-24–3p-siRNA (miR-24–3p siRNA) (n = 26) or negative control-siRNA (control siRNA) (n = 6) for 24 h. Viability (%) was determined using trypan-blue staining normalized to naïve cells (naive). Mean values ± SEM. ** p<0.01.; *** p<0.01. hspa5 (I) and chop (J) expression in naïve RPMI-8226 cells (0 h) normalized to beta-actin and after stimulation with 0,25 μg/ml tunicamycin for 4, 8, and 24h. The relative expression level has been calculated by the __ cT method. Mean values ± SEM. n = 4.

Figure S5. Erk1/2-dependent expression of miR-24–3p. (A) Representative control qPCR of mapk1 expression normalized to beta-actin from RPMI-8226 cells after transfection with either negative control siRNA (control siRNA) or mapk1-specific siRNA (mapk1 siRNA). Relative expression levels were calculated by the __ cT method with negative control as reference (set as 1). (B) c9orf3 expression in RPMI- 8226 cells normalized to beta-actin. RPMI-8226 cells were stimulated with 10 ng/ml, 50 ng/ml and 100 ng/ml of rh-IL-6 for 24 h n = 9. Quantitative analysis of miR-24–3p expression normalized to RNU1–1 in JK-6L (C) and U266 cells (D) treated with 50 ng/ml rh-IL-6 (IL-6); 50 ng/ml rh-IL-6 together with 500 ng/ml of the anti-Erk1/2 peptide (IL-6+anti-Erk1/2) or 10 μM U0126 (IL-6+U0126) for 24 h. n = 9. Quantitative analysis of bim expression normalized to beta-actin in JK-6L (E) and U266 cells (F) treated with 50 ng/ml rh-IL-6 (IL-6); 50 ng/ml rh-IL-6 together with 500 ng/ml of the anti-Erk1/2 peptide (IL-6+anti-Erk1/2) or 10 μM U0126 (IL-6+U0126) for 24 h. n = 9. Quantitative analysis of c9orf3 expression normalized to beta-actin in JK-6L (G) and U266 cells (H) treated with 50 ng/ml rh-IL-6 (IL-6); 50 ng/ml rh-IL-6 together with 500 ng/ml of the anti-Erk1/2 peptide (IL-6+anti-Erk1/2) or 10 μM U0126 (IL-6+U0126) for 24 h. n = 9, n = 3. Relative expression levels were calculated by the __ cT method with naïve cells as reference. (I) RPMI-8226 cells treated with 50 ng/ml rh-IL-6 together with either 10 μM U0126 or 500 ng anti-Erk1/2 peptide for 24 h. Western Blot were performed for pErk1/2 (phospho p44/p42) detection. Beta-actin was used as loading control, the figure is cropped to show pErk lanes. Quantitative analysis of miR-24–3p expression normalized to RNU1–1 and c9orf3 expression normalized to beta actin in JK-6L (J), (L) and U266 cells (K), (M) treated with 100 nmol of specific anti-Erk1/2 (Erk1/2 siRNA) or negative control siRNA (control siRNA) for 48 h. n = 3. The relative expression levels were calculated by the __ cT method. Mean ± SEM. * p<0.05, ** p<0.01, *** p<0.01.

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