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. 2018 Nov 26;7:e38069. doi: 10.7554/eLife.38069

Figure 3. Tsp4b localization to VMS and tenocyte projections requires mechanical force.

Lateral views of live control (A–C) and αBtx injected (D–F) Tg(scx:mCherry) embryos (48 hpf), injected with tsp4b-gfp mRNA showing localization of Tsp4b-GFP (green) (arrowheads) along the VMS and tenocyte projections (red). (I) Histogram shows the percentage of embryos with Tsp4b-GFP localized to VMS (n = 27, p value calculated by chi-squared test <0.05). (G–H) Lateral views of immunostained embryos showing Tsp4b protein localization detected immunohistochemically along VMS in control (G) and αBtx injected (H) embryos. (J) Dot plot shows individual data points of the fluorescent intensity of localized Tsp4b along the VMS in control and αBtx injected embryos. Three VMSs/embryo were sampled in control and αBtx-injected embryos. (n = 9, p value calculated by Wilcoxon Rank Sum Test - < 0.0001). Scale bars = 20 microns. The measurements used for quantitative analysis and creation of the plots can be accessed from Figure 3—source data 1 and Figure 3—source data 2.

Figure 3—source data 1. Count of embryos showing localized or diffuse Tsp4b-GFP.
elife-38069-fig3-data1.csv (178B, csv)
DOI: 10.7554/eLife.38069.023
Figure 3—source data 2. Measurements of Tsp4b fluorescence intensities along VMS.
elife-38069-fig3-data2.csv (1KB, csv)
DOI: 10.7554/eLife.38069.024

Figure 3.

Figure 3—figure supplement 1. Early Lam and Fn organization do not depend on mechanical force.

Figure 3—figure supplement 1.

Dot plot shows individual data points of the fluorescent intensity of localized Lam (A) and Fn (B) at MTJs along the VMS in control and αBtx-injected embryos at 48 hpf (A) and 24 hpf (B). Three VMSs/embryo were sampled in control and αBtx-injected embryos. (n = 9, p value calculated by ANOVA 1-way analysis and Tukey -*<0.002). Scale bars = 20 microns. The measurements used for quantitative analysis and creation of the plots can be accessed from Figure 3—figure supplement 1—source data 1 and Figure 3—figure supplement 1—source data 2.
Figure 3—figure supplement 1—source data 1. Measurement of Laminin fluoresence intensity along VMS.
elife-38069-fig3-figsupp1-data1.csv (1.1KB, csv)
DOI: 10.7554/eLife.38069.018
Figure 3—figure supplement 1—source data 2. Measurement of Fibronectin fluoresence intensity along VMS.
elife-38069-fig3-figsupp1-data2.csv (1.1KB, csv)
DOI: 10.7554/eLife.38069.019
Figure 3—figure supplement 2. Tsp4b organization requires mechanical force.

Figure 3—figure supplement 2.

Lateral view of immunostained 48 hpf embryos showing the localization of Tsp4b in control embryos without (A) and with stimulation (C), and αBtx-injected embryos without (B) and with stimulation (D). Histogram shows the mean area of Tsp4b localization in VMS (dotted region) (E). NS – Not Stimulated, S – Stimulated. Scale bar = 20 microns. The measurements used for quantitative analysis and creation of the plots can be accessed from Figure 3—figure supplement 2—source data 1.
Figure 3—figure supplement 2—source data 1. Measurements of Tsp4b localization area.
elife-38069-fig3-figsupp2-data1.csv (907B, csv)
DOI: 10.7554/eLife.38069.021
Figure 3—figure supplement 3. Mechanical force regulates expression of Tsp4b.

Figure 3—figure supplement 3.

Histogram shows relative expression of rpl13a, scxa and tsp4b in 24 hpf and 48 hpf control and αBtx-injected embryos. (p value calculated by ANOVA 1-way analysis and Tukey test -**<0.001).