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. 2020 Jul 27;39(17):e105696. doi: 10.15252/embj.2020105696

Figure 7. FGF signaling controls ER‐phagy in vivo .

Figure 7

  • A, B
    Representative images of Alcian Blue (cartilage) and Alizarin Red (bone) skeletal staining showing growth retardation in Fgfr3/4 dKO mice compared to age/sex wild‐type littermate at post‐natal day 30. (B) Femur, tibia, and tail details.
  • C
    Hematoxylin/eosin staining of femoral growth plate sections from wild‐type and Fgfr3/4 dKO mice. Higher magnification insets showed a disorganized hypertrophic chondrocyte layer, in Fgfr3/4 dKO mice. Scale bar 60 μm.
  • D
    Western blot analysis of IRS1, p‐P70S6K (T389), P70S6K, p‐JNK (Y185), JNK1/2 proteins in growth plate lysates of mice with indicated genotypes. N = 3 mice/genotype. β‐actin was used as a loading control. Bar graph shows quantification. Mean ± standard error of the mean (SEM). Student's unpaired t‐test *P < 0.05.
  • E
    Western blot analysis of LC3 and Lamp1 proteins from growth plate lysates of mice with indicated genotypes. β‐actin was used as a loading control. N = 3 mice/genotype were analyzed. Bar graph shows quantification. Mean ± standard error of the mean (SEM). Student's unpaired t‐test *P < 0.05.
  • F
    qRT–PCR analysis of Fam134b‐2 expression from growth plate of mice with indicated genotypes. N = 8 (wt mice) and N = 9 (Fgfr3/4 dKO mice) were analyzed. Values were normalized to Hprt gene and expressed as fold change relative to control. Mean ± standard error of the mean (SEM). Student's unpaired t‐test *P < 0.05.
  • G
    Western blot analysis of Fam134b‐2 protein from growth plate lysates of mice with indicated genotypes. β‐actin was used as a loading control. N = 6 (wt mice) and N = 4 (Fgfr3/4 dKO mice) were analyzed. Bar graph showed quantification of Fam134b‐2 normalized to β‐actin. Mean ± standard error of the mean (SEM). Student's unpaired t‐test *P < 0.05.
  • H
    Representative immunofluorescence staining of CLIMP‐63 of femur growth plate sections from mice with indicated genotypes. Scale bar 10 μm. Insets showed increased CLIMP‐63 staining in Fgfr3/4 dKO mice. Scale bar 5 μm. Hypert = hypertrophic chondrocytes; pre‐hypert = pre‐hypertrophic chondrocytes; prolif = proliferating chondrocytes.
  • I
    Representative TEM images of growth plate chondrocytes from mice with indicated genotypes. Arrows indicated the ER. N = nucleus. Scale bar 1 μm.

Source data are available online for this figure.