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. 2016 Apr 18;126(5):1926–1938. doi: 10.1172/JCI83551

Figure 4. Epo induction in Foxd1-Phd2–/– mutants is submaximal.

Figure 4

Shown are the results of multiplex ISH for Epo and EGFP RNA using formalin-fixed, paraffin-embedded kidney tissue sections from Foxd1-mT/mG (Co) and Foxd1-mT/mG-Phd2–/– (Phd2–/–) mutant mice at baseline and after phlebotomy (n = 5 for each group). (A) Representative images of kidney tissue sections containing peritubular interstitial cells expressing Epo (red signal) and/or EGFP (green signal). Nuclei are stained with DAPI (blue signal). Scale bars: 100 μm (baseline and phlebotomy), 10 μm (high-magnification images). White arrows identify Epo-expressing cells; asterisks depict glomeruli. (B) Top left panel: Quantification of Epo-expressing cells. Shown is the absolute number of Epo+ cells per square millimeter of kidney tissue. Bottom left panel: Epo mRNA levels relative to 18S in total kidney tissue homogenates as determined by real-time PCR (n = 3–5). Top right panel: Quantification of EGFP-expressing cells. Shown is the absolute number of EGFP+ cells per square millimeter. Bottom right panel: Fraction of Epo-expressing cells among EGFP+ cells. Data are represented as mean ± SEM; 2-way ANOVA with post hoc Tukey’s test; *P < 0.05, **P < 0.01, ***P < 0.001.