Journal: Frontiers in Endocrinology

Article Title: Panx1 and P2X7R are associated with impaired skeletal health and delayed bone development in early onset of type 1 diabetes

doi: 10.3389/fendo.2026.1829463

Figure Lengend Snippet: Protein expression levels of Panx1 and P2X7R are significantly lower in young adult T1D Akita mice. Protein expression of (A) Panx1 and (B) P2X7R in wild-type (Wt) and T1D Akita mice at 8, 10, and 12 weeks of age. Representative Western blots for Panx1 and P2X7R are shown below the graph. Protein levels for all samples were normalized with the respective loading control, β-actin. P values for comparisons of Akita at each time point with respect to a 8W Wt, b 10W Wt, and c 12W Wt were obtained using one-way ANOVA followed by Tukey’s multiple comparison test. n = 6 (8 weeks), n= 6–8 (10 weeks), n = 7–9 (12 weeks). Data are presented as the means ± SD.

Article Snippet: The membranes were probed with primary polyclonal antibodies to P2X7R (1:1,000; APR-004, Alomone Labs, Israel), Panx1 (N [Term], 1:100; Cat 487900, Invitrogen Corporation, CA), and β-actin (1:20,000; A1978, Sigma-Aldrich) followed by incubation with respective horseradish peroxidase (HP)-conjugated anti-rabbit IgG and anti-mouse IgG (1:10,000; Santa Cruz Biotechnology, TX).

Techniques: Expressing, Western Blot, Control, Comparison

Journal: Frontiers in Endocrinology

Article Title: Panx1 and P2X7R are associated with impaired skeletal health and delayed bone development in early onset of type 1 diabetes

doi: 10.3389/fendo.2026.1829463

Figure Lengend Snippet: Panx1-P2X7R mechanosignaling complex responds differently to mechanical loading in wild-type (Wt) and T1D Akita mice. Relative protein expression of (A) Panx1 and (B) P2X7R in loaded (L) Wt and Akita mice compared to their age-matched non-loaded (nL) Wt after 1, 2 and 4 weeks of loading (treadmill running). Protein levels were first normalized with respective loading control, β-actin, followed by respective age-matched non-loaded controls (denoted as dotted lines) for Wt (blue bars) and Akita (red bars) or respective age-matched loaded Wt (green bars, denoted as dotted line). P values were obtained using an unpaired t-test followed by either Welch’s or Mann-Whitney test. Loading duration groups: n = 6 (1 week), n = 6 (2 weeks), n = 7 (4 weeks). Data presented as means ± SD.

Article Snippet: The membranes were probed with primary polyclonal antibodies to P2X7R (1:1,000; APR-004, Alomone Labs, Israel), Panx1 (N [Term], 1:100; Cat 487900, Invitrogen Corporation, CA), and β-actin (1:20,000; A1978, Sigma-Aldrich) followed by incubation with respective horseradish peroxidase (HP)-conjugated anti-rabbit IgG and anti-mouse IgG (1:10,000; Santa Cruz Biotechnology, TX).

Techniques: Expressing, Control, MANN-WHITNEY

Journal: Frontiers in Endocrinology

Article Title: Panx1 and P2X7R are associated with impaired skeletal health and delayed bone development in early onset of type 1 diabetes

doi: 10.3389/fendo.2026.1829463

Figure Lengend Snippet: Mechanical loading induces different Panx1 and P2X7R transcriptional responses in wild-type (Wt) and Akita mice. Relative mRNA expression of Panx1 ( Panx1 ) (top panel) and P2X7R ( P2rx7 ) (lower panel). (A, D) Non-loaded (nL) Akita mice at 9, 10, and 12 weeks of age compared with age-matched Wt controls. (B, E) Akita and Wt mice after 1, 2, and 4 weeks of treadmill loading (L), comparing loaded versus respective age-matched non-loaded controls. (C, F) Loaded Akita mice compared with their respective age-matched loaded Wt. The delta-delta CT method was used for data analysis, where the value of each gene of interest is first normalized to the reference gene (18S) and then to either their age matched non-loaded Wt (dotted red bars, denoted as dotted lines) or their respective age matched non-loaded controls (denoted as dotted lines) for Wt (blue bars) and Akita (solid red bars) or respective age-matched loaded Wt (green bars). P values for comparisons between Wt nL vs. Akita nL, Wt L vs. Wt nL, Akita L vs. Akita nL, and Akita L vs. Wt L at each time point were obtained using an unpaired t-test followed by either Welch’s or Mann-Whitney test. Loading duration groups: n= 6 (1, 2 weeks), n = 5-7 (4 weeks). Data presented as means ± SD.

Article Snippet: The membranes were probed with primary polyclonal antibodies to P2X7R (1:1,000; APR-004, Alomone Labs, Israel), Panx1 (N [Term], 1:100; Cat 487900, Invitrogen Corporation, CA), and β-actin (1:20,000; A1978, Sigma-Aldrich) followed by incubation with respective horseradish peroxidase (HP)-conjugated anti-rabbit IgG and anti-mouse IgG (1:10,000; Santa Cruz Biotechnology, TX).

Techniques: Expressing, MANN-WHITNEY

Journal: Frontiers in Endocrinology

Article Title: Panx1 and P2X7R are associated with impaired skeletal health and delayed bone development in early onset of type 1 diabetes

doi: 10.3389/fendo.2026.1829463

Figure Lengend Snippet: Exposure to high glucose blunts oscillatory shear stress (OFSS)-induced Panx1 and P2X7R mRNA expression in MLO-Y4 cells. (A) Experimental scheme showing the duration of normal glucose (NG), high glucose (HG), and mannitol (Man) exposure, indicating the points at which 1, 3, and 5 days of 30 minutes OFSS in MLO-Y4 cells were initiated, and sample collection at day 10. Relative mRNA expression of Panx1 ( Panx1 ) and P2X7R ( P2rx7 ) under (B, D) static conditions and (C, E) after 1, 3, and 5 days of OFSS in MLO-Y4 cells conditioned under NG, HG, and mannitol. The delta-delta CT method was used for data analysis, where the value of each gene of interest is first normalized to the reference gene (18S) and then to static NG (B, D) or respective static NG, HG, or Man (C, E) . P values for comparisons among various conditions were obtained using one-way ANOVA followed by Dunnett’s multiple comparisons test. n= 4/condition. Data presented as means ± SD.

Article Snippet: The membranes were probed with primary polyclonal antibodies to P2X7R (1:1,000; APR-004, Alomone Labs, Israel), Panx1 (N [Term], 1:100; Cat 487900, Invitrogen Corporation, CA), and β-actin (1:20,000; A1978, Sigma-Aldrich) followed by incubation with respective horseradish peroxidase (HP)-conjugated anti-rabbit IgG and anti-mouse IgG (1:10,000; Santa Cruz Biotechnology, TX).

Techniques: Shear, Expressing

Journal: Frontiers in Endocrinology

Article Title: Panx1 and P2X7R are associated with impaired skeletal health and delayed bone development in early onset of type 1 diabetes

doi: 10.3389/fendo.2026.1829463

Figure Lengend Snippet: Summary of events driving the load-induced adaptive response in young adult T1D bone compared to healthy bone. The Panx1-P2X7R-mediated mechanosignaling in healthy bone (blue arrows: load adaptation pathways and green arrows: fully adapted pathways) shows a synchronous and adaptive response leading to a robust load-induced anabolic response, while in diabetic bone (red arrows), a dysregulated response together with activation of inflammation and possible recurrence of inflammation that leads to an impaired load-induced anabolic response. Arrows within the boxes represent transcriptional and protein-level changes at each time point.

Article Snippet: The membranes were probed with primary polyclonal antibodies to P2X7R (1:1,000; APR-004, Alomone Labs, Israel), Panx1 (N [Term], 1:100; Cat 487900, Invitrogen Corporation, CA), and β-actin (1:20,000; A1978, Sigma-Aldrich) followed by incubation with respective horseradish peroxidase (HP)-conjugated anti-rabbit IgG and anti-mouse IgG (1:10,000; Santa Cruz Biotechnology, TX).

Techniques: Activation Assay