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. 2020 Dec 28;40(4):e105120. doi: 10.15252/embj.2020105120

Figure 7. Excessive autophagy contributes to renal cystogenesis in OFD type I syndrome.

Figure 7

  1. Western blot of LC3B in kidney homogenates from leupeptin‐treated (40 mg/kg, 6 h) fasted Ofd1 fl/y;creKsp and control mice. Right. LC3B‐II protein levels normalized to Actin (loading control) are expressed as fold increase vs. control mice, n = 4 mice/group.
  2. Representative images of GFP–LC3 puncta (autophagosomes, green) in aquaporin2‐positive (AQP2, red) renal distal tubules from P8 Ofd1fl/y;creKsp;GFP‐LC3 and Ofd1fl/ y;GFP‐LC3 mice. Scale bar = 10 μm. Hoechst labels nuclei. Bar graphs on the left show quantitative analysis of GFP‐LC3 puncta; n = 4 mice/group for a total of 300 AQP2‐positive nuclei/group analyzed.
  3. Western blot of LC3B in kidney homogenates from leupeptin‐treated (40 mg/kg, 6 h) fasted Ofd1‐IND and control mice. Right. LC3B‐II protein levels relative to βTubulin (loading control) are expressed as fold increase compared with control mice, n = 5 mutant/8 control mice.
  4. Western blot of Atg13 in Ofd1 fl/y;creKsp and control kidneys at pre‐cystic (P8, left) and cystic (P22, right) stages. Gapdh was used as loading control. Atg13 protein levels are expressed as fold change compared with control mice; n = 7 mutant/5 control mice at P8, n = 4 mice/group at P22.
  5. Hematoxylin and eosin staining of kidney sections from Ofd1 fl/y;Atg7+/+;creKsp and Ofd1 fl/y;Atg7 fl/fl;creKsp mice. Right. Graph shows fold change of the cysts number observed in Ofd1 fl/y;Atg7 fl/fl;creKsp animals compared with Ofd1 fl/y;Atg7+/+;creKsp; n = 12 sections/animal were analyzed; n = 6 mice/group. Scale bars = 1 mm.
  6. Blood Urea Nitrogen was quantified on blood withdrawn from Ofd1 fl/y;Atg7 fl/fl;creKsp, Ofd1 fl/y;Atg7+/+;creKsp and control mice at P45. n = 8 mice per group.

Data information: Data are expressed as mean values from the indicated independent experiments. Error bars indicate SEM. One‐tailed Student’s t‐test was applied; *P ≤ 0.05, **P ≤ 0.01, and ***P ≤ 0.001. BUN = blood urea nitrogen.

Source data are available online for this figure.