Supplementary_Table_3 – Supplemental material for Contributors, risk associates, and complications of frailty in patients with chronic kidney disease: a scoping review

Supplemental material, Supplementary_Table_3 for Contributors, risk associates, and complications of frailty in patients with chronic kidney disease: a scoping review by Patrick Yihong Wu, Chia-Ter Chao, Ding-Cheng Chan, Jenq-Wen Huang and Kuan-Yu Hung in Therapeutic Advances in Chronic Disease</p

Additional file 2 of The down-regulation of XBP1, an unfolded protein response effector, promotes acute kidney injury to chronic kidney disease transition

Additional file 2: Figure S1. UIRI causes prominent renal damage and development of fibrosis. (a) Diagram illustrates the timeline of the experiment. The left kidney of male C57BL/6 mice was subjected to renal ischemia/reperfusion injury (UIRI) and then sacrificed at different days as indicated. UDx: x days after UIRI. (b) PAS staining represents the accumulation of debris in the tubular lumen after UIRI. The arrowhead in the lower panel indicates debris. Scale bar indicates 200 μm in 40x, 50 μm in 200x. (c) qPCR assessment of the relative expression level of Kim-1 mRNA. (d) Masson’s trichrome staining shows the increased fibrosis fraction in kidney section after UIRI. Scale bar indicates 50 μm in 200x. (e) Quantitative scores of interstitial fibrosis were assessed. (f and g) The expression of α-SMA was examined with western blot analysis and quantified. Data are expressed as means ± SEM, n = 3 ~ 6 in each group. * P < 0.05 and *** P < 0.001, as compared with sham group. Figure S2. Loss of XBP1 expression is a universal characteristic in renal fibrosis models. (a) Western blot analysis showed the protein expression of α-SMA, XBP1u and XBP1s in UUO mice model. GAPDH was used as an internal control. (b-d) Quantification of relative protein expression levels of α-SMA, XBP1u and XBP1s. (e) Western blot analysis showed the protein expression of α-SMA, XBP1u and XBP1s in adenine diet mice model. GAPDH was used as an internal control. (f–h) Quantification of relative protein expression levels of α-SMA, XBP1u and XBP1s. N = 3–4 for each group, * P < 0.05, ** P < 0.01, and *** P < 0.001, as compared with sham or chow diet group. Figure S3. Proximal tubular conditional knockout mice blocked XBP1s activation. (a) Diagram illustrates SLC5aCreERT2; XBP1fl/fl mice. (b) After tamoxifen administration, mice were subjected to IP injection of 500 ng/g of Tunicamycin for 12 h. Western blot analysis showed protein expression of XBP1s after Tunicamycin induction in XBP1fl/fl or XBP1cKO mice. GAPDH was used as an internal control. (c) Immunofluorescence staining demonstrated XBP1 expression in Tunicamycin treated mice kidneys. Scale bar: 250 μm. (d) XBP1s mRNA expression level was determined by semi-quantitative PCR. (e) qPCR assessment of the relative expression level of XBP1s mRNA. Figure S4. Proximal tubular XBP1 specific knockout mice were vulnerable to UIRI-induced kidney injury. (a) Diagram illustrates the experimental timeline of tamoxifen administration and UIRI with contralateral nephrectomy (Nx) surgery in XBP1fl/fl and XBP1cKO mice. (b and c) Blood urea nitrogen (BUN) and serum creatinine (Scr) levels were measured after 1 day of contralateral Nx. N = 3 for each group. ** P < 0.01, and *** P < 0.001, as compared with XBP1fl/fl Sham group. Figure S5. UIRI induces cell cycle arrest in G2/M phase. (a-d) The expression of chk1 and p21 in mice kidneys was evaluated with western blotting and quantified. GAPDH was used as an internal control. Data are expressed as means ± SEM, n = 3 ~ 6 in each group. * P < 0.05, ** P < 0.01, and *** P < 0.001, as compared with sham group. (e and f) Representative images of Ki67+ pHH3+ renal sections in (e) WT mice or (f) XBP1cKO and XBP1fl/fl mice subjected to UIRI or Sham operation. Selected areas indicated highly expressed double-positive tubules. Scale bar: 50 μm. (g) Number of Ki67+pHH3+ tubular cells. Data are expressed as means ± SEM. * P < 0.05, ** P < 0.01, and *** P < 0.001, as compared with XBP1fl/fl sham group. ### P < 0.001 compared between indicated groups

Additional file 3 of The down-regulation of XBP1, an unfolded protein response effector, promotes acute kidney injury to chronic kidney disease transition

Additional file 3. Proteomic analysis of differentially expressed proteins in HK-2 cells silenced of XBP1 compared to scramble control

Multivariable logistic regression models for parameters associated with left ventricular diastolic dysfunction.

<p>Models 2–4 adjust for age, gender, BMI, DM, HTN, HOMA, and LDL. Models 5 and 6 additionally adjust for interleukin-6.</p><p>Abbreviations: BMI, body mass index; HOMA, homeostasis model of insulin resistance; DM, diabetes mellitus; HTN, hypertension; LV, left ventricular; IL-6, interleukin-6; LDL, low-density lipoprotein.</p

The designed patient flow diagram.

<p>The designed patient flow diagram.</p

Correlation between interleukin-6 (IL-6), tissue necrosis factor-alpha (TNF-α), and echocardiographic diastolic function parameters in PD patients.

<p>(A) TNF-α and mitral valve ejection flow deceleration time (DT, r = 0.58, P<0.001); (B) TNF-α and the ratio of mitral valve ejection flow (E) divided by mitral valve atrium flow (A, r = −0.38, P<0.001); (C) TNF-α and the ratio of E divided by early diastolic lengthening velocities in tissue Doppler imaging (Em, r = 0.62, P<0.001); (D) IL-6 and DT (r = 0.71, P<0.001); (E) IL-6 and E/A (r = −0.49, P<0.001); and (F) IL-6 and E/Em (r = 0.62, P<0.001).</p

The prevalence of left ventricular (LV) diastolic dysfunction in PD patients divided into tertiles based on interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) levels.

<p>The prevalence of left ventricular (LV) diastolic dysfunction in PD patients divided into tertiles based on interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) levels.</p

Western blots of integrin-activated kinases.

<p>Equal amounts of protein (20 µg per lane) from untreated or fibrin-covered PMCs were resolved, transferred, and probed for phosphorylated FAK (focal adhesion kinase) and Src (A). The relative levels of Src/GAPDH (B) and FAK/GAPDH (C) were measured by densitometry. C, PMCs without fibrin; F10, fibrin covered for 10 min; P10, fibrin covered and treated with pentoxifylline 0.3 mg/ml for 10 min; A10, fibrin covered and treated with α_vβ₃ integrin antibody for 10 min; F20, fibrin covered for 20 min; F30, fibrin covered for 30 min. *p<0.05 vs. Control.</p

Additional file 1 of The down-regulation of XBP1, an unfolded protein response effector, promotes acute kidney injury to chronic kidney disease transition

Additional file 1: Table S1. Primer sequences of PCR, real-time PCR, and genotyping

Changes in cell markers after application of fibrin to peritoneal mesothelial cells (PMCs).

<p>Expression of α-smooth muscle actin (α-SMA), fibronectin, fibroblast specific protein-1 (FSP-1), and β₃ integrin were detected by immunofluorescence staining with FITC-labeled secondary antibodies (green). Nuclei were counterstained with PI (red). PMCs overlaid with fibrin for 4 h (+Fibrin) expressed higher levels of α-SMA, fibronectin, FSP-1, and β₃ integrin than untreated PMCs (-Fibrin). Original magnification ×400.</p