Fig 1: Expression of caspase-12 in osteoblasts. Comparison of serial sections of the mouse mandibular bone at the prenatal/embryonic (E) day 15 showing co-localization of caspase-12 (A) with osteoblastic markers osteocalcin (B) and Runx2 (C) within the bone. Positive cells are brown, counterstained by hematoxylin (blue). Arrows point to cells in the same region. Scale bar: 50 μm. Expression of caspase-12 in undifferentiated and differentiated MC3T3-E1 cells (D) showing stable expression of Casp12 after 21 days of differentiation. Western blot analysis (E) and immunofluorescence (F) confirmed the presence of caspase-12 protein in MC3T3-E1. E: 55 kDa-full length caspase-12, 42 kDa-cleaved caspase-12, DMSO: caspase-12 forms after DMSO treatment, C12 INH: caspase-12 forms after specific inhibitor treatment. F: Caspase-12 in green, DAPI in blue. Scale bar: 100 μm.
Fig 2: Gene silencing of Casp12 in MC3T3-E1. Gene silencing of Casp12 in MC3T3-E1 cells using specific siRNA showed statistically significant reduction of Casp12 expression (A). Real-time PCR confirmed statistically significant decrease in expression of Alpl (D), Bglap (E), Phex (F), and Dmp1 (G) after Casp12 silencing. Results are presented as means ± standard deviations and were analyzed using two-tailed t-test. Immunofluorescence detection of caspase-12 (green) protein in MC3T3-E1 after Casp12 gene silencing (B) compared to control (C), DAPI in blue.
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