Fig. 4: T2D-induced specific reduction of SERCA2 expression mediates an attenuation of osteocyte mechano-responsiveness. | Nature Communications

Fig. 4: T2D-induced specific reduction of SERCA2 expression mediates an attenuation of osteocyte mechano-responsiveness.

From: Rescuing SERCA2 pump deficiency improves bone mechano-responsiveness in type 2 diabetes by shaping osteocyte calcium dynamics

Fig. 4

a Real-time intracellular Ca2+ signaling of osteocytes in situ in mouse tibiae treated with DMSO (vehicle control) and two antagonists of SERCA, i.e., thapsigargin (1 μM) and cyclopiazonic acid (CPA; 5 μM), in response to subsequent cyclic mechanical loading. b Intracellular Ca2+ signaling in MLO-Y4 osteocytic cells in vitro treated with DMSO, thapsigargin, and CPA in response to subsequent steady FSS stimulation at 2 Pa. c qRT-PCR analyses of the ATP2A1, ATP2A2 and ATP2A3 gene expression in MLO-Y4 osteocytic cells treated with the normal medium (NC), mannitol-supplemented medium (MAN, osmotic pressure-matched control), and high-glucose and high-fat (HGHF)-supplemented medium. d Western blotting analyses of the SERCA1, SERCA2, and SERCA3 protein expression in normal, mannitol-treated, and HGHF-treated MLO-Y4 osteocytic cells. e qRT-PCR analyses of the ATP2A1, ATP2A2 and ATP2A3 gene expression in male non-diabetic, KK-Ay, and HFD + STZ mice. f Immunohistochemical staining of SERCA1, SERCA2, and SERCA3 in osteocytes in the mouse tibiae of the non-diabetes, KK-Ay, and HFD + STZ groups. g Intracellular Ca2+ signaling in MLO-Y4 osteocytic cells in vitro treated with siCtrl and siSERCA2 in response to steady FSS stimulation at 2 Pa. Graphs represent mean ± SD (a, b, g: n = 120 cells per group; c–e n = 6 biologically independent replicates; f: n = 8 mice per group). ***P < 0.001 by one-way ANOVA with Bonferroni’s post test. Specific P values are provided in the Source Data file. Scale bars: f 20 μm.

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