Figure 8

Advanced fibrotic injuries cause renal developmental program reactivation in the distal nephron tubular segments of adult kidney. (a) Dot plot of cell type-specific expression of Hoxb6, Cd24a and Sox4 for manually annotated clusters in the control, UIR and UUO kidney. Dot size denotes percentage of cells expressing the marker. Color intensity represents average gene expression values. (b) Representative RNAscope images for Hoxb6 (green), Lrp2 (magenta), Slc12a3 (white) and DAPI in the control and UIR kidneys. Original magnification, × 60, maximal intensity projection, 0.14 μm/px zoom. White arrows show Hoxb6 expression in Lrp2-nehative and Slc12a3-positive tubules. (c) Sox4 and Cd24a qPCR in control and fibrotic kidneys, n = 4–7 per group. (d) Representative bands and quantifications of Sox4 and Cd24a Western blots, n = 4–5 per group. Representative bands are cropped out of the original gels and are separated by the black border, the unprocessed original blots/gels are presented in Supplementary Fig. S29. *P ≤ 0.05, ***P ≤ 0.001, ****P ≤ 0.0001 compared to control, Student’s t test for (c) and (d). (e) Representative images of combined IF for Sox4 (green), Umod (yellow), DAB (magenta), LTL (white) and DAPI (blue), control and UIR kidneys. Original magnification, maximal intensity projection, × 60, 0.09 μm/px zoom. Areas of Sox4 co-localization with Umod are shown with white arrows.