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Fig. 6 Mechanical force regulates expression of klf2a in mhj myotendinous junction tenocytes. Ventral views of mandibulohyoid junction (mhj), myotendinous junction (MTJ) associated tenocytes showing in situ Hybridization Chain Reaction (isHCR) of klf2a (green) and anti-mCherry immunofluorescence (magenta) marking the tenocytes in Tg(scxa:mCherry) embryos at 72 hpf in WT uninjected (WT) (A–E), aBTX-inj (Paralyzed) (F–J), partially recovered aBTX-inj (Twitching) (K–O), and completely recovered aBTX-inj (Full Recovery) (P–T) conditions. (D, I, N, S) Grayscale images showing nuclei stained with DAPI with ROIs showing isolated 3D volumes of mhj tenocytes (magenta) based on DAPI signal. (E, J, O, T) Insets showing magnified views of the 3D volumes of tenocytes associated with mhj MTJ depicting expression of klf2a and stained for mCherry. (U) Violin plot showing changes in mean fluorescence intensity of klf2a in mhj MTJ tenocyte nuclei between WT (n = 17), Paralyzed (n = 15), Twitching (n = 14), and Full Recovery (n = 11) with ~10 nuclei measured per embryo. p-value calculated with linear mixed effects model with Tukey post hoc test. *p < 0.05, ***p < 0.001. Scale bars = 20 µm.