Fig. 5

Evaluation of permselectivity using human membranous nephropathy serum samples. a Representation of the GOAC albumin permselectivity assay to MN serum exposure. Following a 24 h incubation with media supplemented with 0.5% serum from healthy individuals (CTRL1 and CTRL2) or MN patients (MN1–6), albumin-FITC is applied to channel C (yellow) and flow-through collected in channel F. b–g Bright field showing albumin leakage after 5 min (left columns) and 60 min (right columns) after exposure to healthy and MN patients serum in hAKPC-P + hGEC (b, c), hiPOD + hGEC (d, e), and hpPOD + hGEC (f, g) chips. Leakage is evident in hAKPC-P + hGEC and hpPOD + hGEC but not in hiPOD + hGEC chips after exposure to MN serum. h–j Box plot graph of fluorescein absorbance (expressed as log) in filtrate collected after 60 min after serum exposure in hAKPC-P + hGEC (h), hiPOD + hGEC (i), and hpPOD + hGEC (j) chips. Statistically significant increase in albumin permeability is evident after exposure to MN sera only in hAKPC-P + hGEC and hpPOD + hGEC chips. Number of replicates for chips used in h is as follows: hAKPC-P + hGEC chip and CTRL1: #7; hAKPC-P + hGEC chip and CTRL2: #8; hAKPC-P + hGEC chip and MN1: #4 ; hAKPC-P + hGEC chip and MN2: #7; hAKPC-P + hGEC chip and MN3: #3. hAKPC-P + hGEC chip and MN4: #4; hAKPC-P + hGEC chip and MN5: #7; hAKPC-P + hGEC chip and MN6: #3. Number of replicates for chips used in i as follow: hiPOD + hGEC chip and CTRL1: #6; hiPOD + hGEC chip and CTRL2: #4; hiPOD + hGEC chip and MN1: #11; hiPOD + hGEC chip and MN2: #5; hiPOD + hGEC chip and MN3: #4; hiPOD + hGEC chip and MN4: #11; hiPOD + hGEC chip and MN5: #5; hiPOD + hGEC chip and MN6: #4. Number of replicates for chips used in j is as follows: hpPOD + hGEC chip and CTRL1: #8; hpPOD + hGEC chip and CTRL2: #9; hpPOD + hGEC chip and MN1: #6; hpPOD + hGEC chip and MN2: #7; hpPOD + hGEC chip and MN3: #7; hpPOD + hGEC chip and MN4: #6; hpPOD + hGEC chip and MN5: #7; hpPOD + hGEC chip and MN6: #7. For d, g, j significant differences were determined by a one-way ANOVA and Holm–Sidak post hoc test, *p < 0.05, **p < 0.01, ***p < 0.001 Box plots show the median, the 25th and 75th percentiles, whiskers (median ± 1.5 times interquartile range), and outliers (solid circle)