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. 2022 Jan 25;298(3):101639. doi: 10.1016/j.jbc.2022.101639

Figure 1.

Figure 1

Deletion of Pip5k1c in limb and head mesenchymal stem cells does not affect skeletal development in mice.A, genotyping. Representative PCR analysis of genomic DNA extracted from tail clippings. PCR product bands of floxed (380 bp) and control (WT) (250 bp) are shown. Cre PCR product band (650 bp) is indicated. B, real-time quantitative reverse transcription PCR (qRT-PCR) analysis. Total RNA isolated from the indicated tissues of E14.5 Pip5k1cPrx1cKO (cKO) or Cre-negative Pip5k1cfl/fl (WT) littermates were subjected to qRT-PCR analysis for Pip5k1c expression. FL, forelimb. Pip5k1c mRNA was normalized to Gapdh mRNA. Results were expressed as mean ± standard deviation (SD); N = 3 mice per group. ∗∗p < 0.01 versus control; Unpaired two-tailed t test. C, gross appearance of P0 control and cKO mice. Scale bar, 1 cm. D, body weight of P0 control and cKO mice. Results were expressed as mean ± SD; N = 5 mice for control, N = 6 mice for cKO. n.s., not significant; Unpaired two-tailed t test. E, alizarin red and alcian blue double staining of P0 control and cKO skeletons. Scale bar, 1 cm. F, calvaria, forelimb, hindlimb, sternum, and clavicle from P0 control and cKO mice. Scale bar, 1 cm. G, quantitative data of full-length (FL) and bony portion length of the indicated bones from (E). Results were expressed as mean ± SD; N = 3 mice per group. Unpaired two-tailed t test. H, alcian blue staining of femur sections from P0 control and cKO mice. Scale bar, 200 μm. I, quantitative data from (H). Results were expressed as mean ± SD; N = 3 mice per group. n.s., not significant; unpaired two-tailed t test. J, H/E staining of P0 control and cKO femur sections. Representative images of proliferation zone (PZ) and hypertrophic zone (HZ) from P0 control and cKO femurs. Scale bar, 100 μm. K, alcian blue staining of P17 control and cKO tibiae. The secondary ossification centers are shown. Scale bar, 100 μm. BP, bony portion; n.s., not significant; PZ, proliferation zone; HZ, hypertrophic zone.