Figure 2. Effect of R246Q mutation on podocyte related gene transcription.

(a) Western blot confirming expression of the viral transgene with the presence of the MYC tag in myc-LMX1B_WT or myc-LMX1B_R246Q infected immortalized human podocyte cells compared to uninfected and lentiviral control podocytes (top). β-actin was used as a loading control for all samples (bottom). Blots were cropped to exclude lanes that are not relevant to this paper, the uncropped blot is displayed in Supplementary Figure 6. (b) Graph depicting average Relative Quantity (RQ) values for NPHS1, GLEPP 1, INF2, and SYNPO in myc-LMX1B_WT or myc-LMX1B_R246Q cells (N ≥ 3). Error bars = SEM *p ≤ 0.05, **p ≤ 0.001 using t-test. (c) Immunoblotting showing reduced translation of nephrin (n = 7, p = 0.027) but not GLEPP1 (n = 4, p > 0.05), CD2AP (n = 3, p > 0.05), synaptopodin (n = 3, p > 0.05) and INF2 (n = 3, p > 0.05) in myc-LMX1B_R246Q podocytes compared to myc-LMX1B_WTexpressing cells. β-actin was used as a loading control for all samples and unedited blots are shown in Supplementary Figure 6.