Fig 1: Panx1 and 2 expression in mouse LG. (A) Panx1 expression slightly decreases in the LG of adult (postnatal day 60 [P60]) mice compared with the LG obtained from mice at postnatal day 1 (P1). (B) Expression of Panx2 increases in adult LGs compared with LGs at P1. In A and B, histograms represent the normalized expression of Panx1 and 2 to GAPDH assessed by qRT-PCR in three independent experiments. (C–H) Panx1 expression in mouse LGs at P0 (C) during postnatal (D: P20) development and adulthood (E–H). At P and P20, Panx1 (green) is highly expressed in the developing LG ducts (C, D). Panx1 expression in the developing acini is low (D). Myoepithelial cells (red) are localized around acini, nuclei labeled with 4′,6-Diamidine-2-phenylindole (DAPI) (blue). (E, F, H) Panx1 expression in adult LG (P60). (F–H) MECs have low level of Panx1 expression. Low level of Panx1 expression in MEC is confirmed in nonmerged images (F, Panx1 expression; green; and G, SMA [marker of MECs] expression, red). (I) Low level of Panx2 expression in the epithelial components of the LG at P. Panx2 was only detected in blood vessels (white arrow) and some mesenchymal cells (yellow arrows) At P60, Panx2 (J, K) is expressed in the acinar cells. High levels of Panx2 protein is found in blood vessels (J, white arrows) and apical parts of acinar cells (K, apical; basal parts of acinar cells are labeled with puncture line). Myoepithelial cells (red). Nuclei are labeled with DAPI (blue).
Fig 2: Panx1 deletion does not alter bone formation or osteoclast number/surface in cortical bone from female mice. (A) Periosteal (Ps) and endocortical (Ec) MS/BS, MAR, and BFR/BS were measured in unstained femoral cross-sections (n = 9–12 per group). (B) N.Oc/BS, Oc.S/BS, ES/BS were scored on the endocortical surface of the femoral mid-diaphysis after staining for TRAPase/Toluidine blue in Panx1fl/fl and Panx1?ot mice at 4 and 13 months of age (n = 9–12 per group). Bars represent mean ± s.d., Lines indicate age effect, by 2-way ANOVA.
Fig 3: Panx1?ot female mice exhibit increased muscle mass. (A–D) Body weight (A), total lean body mass (B), percent of lean and fat mass (C), and BMD (D) were measured in 10-week-old female and male mice by DXA (Piximus) (n = 9–13 per group). (E) mRNA and protein Panx1 levels were measured in tibialis anterior (TA) muscles by qPCR (corrected by GAPDH, n = 9 and 12 per group), and Western blotting (corrected by ß-actin, n = 4 and 3), respectively. Bars represent mean ± s.d., *p < 0.05 versus Panx1fl/fl mice of the same sex, by 2-way ANOVA.
Fig 4: Deletion of Panx1 in osteocytes increases bone mass. (A) Panx1 protein levels in calvaria bones from Panx1fl/fl and Panx1?ot mice at 4 months of age were assessed by Western blotting and ß-actin was used as a loading control. (B) Representative images of whole body histological preparations (n = 3/genotype) stained by Alcian blue/alizarin red in 6-day-old mice for visual inspection of cartilage (blue) and calcified tissue (magenta). (C) The levels of the apoptosis-related genes were measured by qPCR bone marrow-flushed long bones obtained from female mice. Bars represent mean ± s.d., #p < 0.05 versus Panx1fl/fl mice at the same age, *p < 0.05 versus 4-month-old mice of the same genotype, and lines indicate age effect, by 2-way ANOVA. (D) Body weight and total body, spinal and femoral BMD were assessed monthly in female mice from 1 to 13 months of age by DXA (n = 9–12 per group). ¥p < 0.05 versus Panx1fl/fl mice at the same age by t-test.
Fig 5: Deletion of Panx1 in osteocytes decreases osteoclastic gene expression in aged 13-month-old female mice. (A) CD11b mRNA levels were measured in non-adherent bone marrow cells by qPCR and corrected by GAPDH (n = 9–12 per group). (B–D) mRNA levels of RANKL and OPG as well as the indicated osteoclast-related genes were measured in bone marrow-flushed long bones by qPCR and corrected by GAPDH (n = 9–12 per group). Bars represent mean ± s.d., #p < 0.05 versus Panx1fl/fl mice at the same age, *p < 0.05 versus 4-month-old mice of the same genotype, and lines indicate age effect, by 2-way ANOVA.
Supplier Page from MilliporeSigma for Anti-PANX1 antibody produced in rabbit