FIGURE

Figure 4

ID
ZDB-FIG-230211-97
Publication
Tonelli et al., 2023 - Zebrafish Tric-b is required for skeletal development and bone cells differentiation
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Figure 4

Collagen type I and ultrastructural analysis. (A) Representative Coomassie stained SDS-Urea-PAGE of collagen type I extracted from WT (n = 2) and mutants (tmem38b-/- n = 2; tmem38bΔ120-7/Δ120-7 n = 2) bone and skin. Mutants’ collagen α bands presented a slight faster migration compared to WT as highlighted by red lines. (B) Transmission electron microscopy images of 1 mpf WT (n = 3), tmem38b-/- (n = 3), and tmem38bΔ120-7/Δ120-7 (n = 3) fibroblasts and osteoblasts at the vertebral endplate. ER cisternae enlargement was evident in mutants (arrowheads). Magnification 80000x. Scale bar: 200 nm. The ER cisternae area quantitation is shown in the graphs (bottom). (C) Immunohistochemistry of 5 dpf WT and tmem38b-/- with Hsp47b antibody. High magnification details of the skin of WT and tmem38b-/- following immunostaining with Hsp47b antibody revaled that Hsp47 signal was located in the fibroblasts of the skin (arrowheads). (D) Representative images of 5 dpf fish after whole mount immunohistochemistry with Hsp47b antibody, before and after 4PBA administration. Scale bar: 500 μm. (E) Analysis of Hsp47b expression by whole mount immunohistochemistry in WT and in tmem38b-/- before and after 4PBA administration. A significant increase of Hsp47 signal was detected in tmem38-/- compared to WT. 4PBA treatment significantly reduced the Hsp47b signal in tmem38b-/- compared to untreated mutant fish without reaching WT value (WT n = 146 and tmem38b-/- n = 116 untreated and 4PBA treated tmem38b-/- n = 113). (F) Analysis of Hsp47b expression by whole mount immunohistochemistry in WT and in tmem38bΔ120-7/Δ120-7 before and after 4PBA administration. A significant increase of Hsp47 signal was present in tmem38bΔ120-7/Δ120-7 compared to WT. 4PBA treatment significantly reduced the Hsp47b signal in tmem38bΔ120-7/Δ120-7 compared to untreated mutant fish reaching WT value (WT n = 43 and tmem38bΔ120-7/Δ120-7 n = 19 untreated and 4PBA treated tmem38bΔ120-7/Δ120-7 n = 34). Zero (nd), low (+) and high (++) indicate the intensity of the signal. *p < 0.05, **p ≤ 0.01, ***p ≤ 0.001.

Expression Data

Expression Detail
Antibody Labeling
Phenotype Data
Fish:
Condition:
Observed In:
Stage Range: Day 5 to Adult

Phenotype Detail
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Front Endocrinol (Lausanne)