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(A) Representative images of SCA-1 immunostaining of longitudinal radius sections at two months focused on the mid-diaphysis of Cas9 eGFP ; Prx1 -cre control and PCC mice (n=4 Cas9 eGFP ; Prx1 -cre and n=6 PCC). (B) Quantification of fibrous layer thickness (μm) at the mid-diaphysis, ROI represented in (A). (C) Representative images for Periostin (POSTN, magenta) and Ki67 (proliferation, green) immunostaining of longitudinal radius sections at two months (n=6 Cas9 eGFP ; Prx1 -cre and n=8 PCC). (D) Quantification of proliferation index (% Ki67+) at the mid-diaphysis, ROIs represented in (C). Significant periosteal expansion results from increased proliferation of cells in the cambium layer. (E) PECAM staining of longitudinal radius sections at two months (n=4 Cas9 eGFP ;Prx1- cre and n=5 PCC). (F) Representative images for RUNX2 (magenta) and Ki67 (green) immunostaining at the mid-diaphysis (n=4 Cas9 eGFP ; Prx1 -cre and n=4 PCC). (G) Quantification of RUNX2+ (magenta bars), Ki67+ (green bars), and RUNX2+Ki67+ double positive cells (magenta and green striped bars) in ROIs represented in (F). (H) Representative images for <t>COL1A1</t> (magenta) and Ki67 (green) immunostaining at the mid-diaphysis (n=6 Cas9 eGFP ; Prx1 -cre and n=6 PCC). (I) Quantification of COL1A1+ (magenta bars), Ki67+ (green bars), and Col1a1+Ki67+ double positive cells (magenta and green striped bars) in ROIs represented in (H). B = bone. c = cambium layer. f = fibrous layer. Ma = marrow. Scale bars = 50 μm. Arrowheads indicate Ki67+ cells. Data represent mean +/-SD. * denotes Welch’s t-test <0.05. ** denotes Welch’s t-test <0.005. *** denotes Welch’s t-test <0.0005. ns = not significant.
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(A) Representative images of SCA-1 immunostaining of longitudinal radius sections at two months focused on the mid-diaphysis of Cas9 eGFP ; Prx1 -cre control and PCC mice (n=4 Cas9 eGFP ; Prx1 -cre and n=6 PCC). (B) Quantification of fibrous layer thickness (μm) at the mid-diaphysis, ROI represented in (A). (C) Representative images for Periostin (POSTN, magenta) and Ki67 (proliferation, green) immunostaining of longitudinal radius sections at two months (n=6 Cas9 eGFP ; Prx1 -cre and n=8 PCC). (D) Quantification of proliferation index (% Ki67+) at the mid-diaphysis, ROIs represented in (C). Significant periosteal expansion results from increased proliferation of cells in the cambium layer. (E) PECAM staining of longitudinal radius sections at two months (n=4 Cas9 eGFP ;Prx1- cre and n=5 PCC). (F) Representative images for RUNX2 (magenta) and Ki67 (green) immunostaining at the mid-diaphysis (n=4 Cas9 eGFP ; Prx1 -cre and n=4 PCC). (G) Quantification of RUNX2+ (magenta bars), Ki67+ (green bars), and RUNX2+Ki67+ double positive cells (magenta and green striped bars) in ROIs represented in (F). (H) Representative images for <t>COL1A1</t> (magenta) and Ki67 (green) immunostaining at the mid-diaphysis (n=6 Cas9 eGFP ; Prx1 -cre and n=6 PCC). (I) Quantification of COL1A1+ (magenta bars), Ki67+ (green bars), and Col1a1+Ki67+ double positive cells (magenta and green striped bars) in ROIs represented in (H). B = bone. c = cambium layer. f = fibrous layer. Ma = marrow. Scale bars = 50 μm. Arrowheads indicate Ki67+ cells. Data represent mean +/-SD. * denotes Welch’s t-test <0.05. ** denotes Welch’s t-test <0.005. *** denotes Welch’s t-test <0.0005. ns = not significant.
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(A) Representative images of SCA-1 immunostaining of longitudinal radius sections at two months focused on the mid-diaphysis of Cas9 eGFP ; Prx1 -cre control and PCC mice (n=4 Cas9 eGFP ; Prx1 -cre and n=6 PCC). (B) Quantification of fibrous layer thickness (μm) at the mid-diaphysis, ROI represented in (A). (C) Representative images for Periostin (POSTN, magenta) and Ki67 (proliferation, green) immunostaining of longitudinal radius sections at two months (n=6 Cas9 eGFP ; Prx1 -cre and n=8 PCC). (D) Quantification of proliferation index (% Ki67+) at the mid-diaphysis, ROIs represented in (C). Significant periosteal expansion results from increased proliferation of cells in the cambium layer. (E) PECAM staining of longitudinal radius sections at two months (n=4 Cas9 eGFP ;Prx1- cre and n=5 PCC). (F) Representative images for RUNX2 (magenta) and Ki67 (green) immunostaining at the mid-diaphysis (n=4 Cas9 eGFP ; Prx1 -cre and n=4 PCC). (G) Quantification of RUNX2+ (magenta bars), Ki67+ (green bars), and RUNX2+Ki67+ double positive cells (magenta and green striped bars) in ROIs represented in (F). (H) Representative images for <t>COL1A1</t> (magenta) and Ki67 (green) immunostaining at the mid-diaphysis (n=6 Cas9 eGFP ; Prx1 -cre and n=6 PCC). (I) Quantification of COL1A1+ (magenta bars), Ki67+ (green bars), and Col1a1+Ki67+ double positive cells (magenta and green striped bars) in ROIs represented in (H). B = bone. c = cambium layer. f = fibrous layer. Ma = marrow. Scale bars = 50 μm. Arrowheads indicate Ki67+ cells. Data represent mean +/-SD. * denotes Welch’s t-test <0.05. ** denotes Welch’s t-test <0.005. *** denotes Welch’s t-test <0.0005. ns = not significant.
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(A) Representative images of SCA-1 immunostaining of longitudinal radius sections at two months focused on the mid-diaphysis of Cas9 eGFP ; Prx1 -cre control and PCC mice (n=4 Cas9 eGFP ; Prx1 -cre and n=6 PCC). (B) Quantification of fibrous layer thickness (μm) at the mid-diaphysis, ROI represented in (A). (C) Representative images for Periostin (POSTN, magenta) and Ki67 (proliferation, green) immunostaining of longitudinal radius sections at two months (n=6 Cas9 eGFP ; Prx1 -cre and n=8 PCC). (D) Quantification of proliferation index (% Ki67+) at the mid-diaphysis, ROIs represented in (C). Significant periosteal expansion results from increased proliferation of cells in the cambium layer. (E) PECAM staining of longitudinal radius sections at two months (n=4 Cas9 eGFP ;Prx1- cre and n=5 PCC). (F) Representative images for RUNX2 (magenta) and Ki67 (green) immunostaining at the mid-diaphysis (n=4 Cas9 eGFP ; Prx1 -cre and n=4 PCC). (G) Quantification of RUNX2+ (magenta bars), Ki67+ (green bars), and RUNX2+Ki67+ double positive cells (magenta and green striped bars) in ROIs represented in (F). (H) Representative images for <t>COL1A1</t> (magenta) and Ki67 (green) immunostaining at the mid-diaphysis (n=6 Cas9 eGFP ; Prx1 -cre and n=6 PCC). (I) Quantification of COL1A1+ (magenta bars), Ki67+ (green bars), and Col1a1+Ki67+ double positive cells (magenta and green striped bars) in ROIs represented in (H). B = bone. c = cambium layer. f = fibrous layer. Ma = marrow. Scale bars = 50 μm. Arrowheads indicate Ki67+ cells. Data represent mean +/-SD. * denotes Welch’s t-test <0.05. ** denotes Welch’s t-test <0.005. *** denotes Welch’s t-test <0.0005. ns = not significant.
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(A) Representative images of SCA-1 immunostaining of longitudinal radius sections at two months focused on the mid-diaphysis of Cas9 eGFP ; Prx1 -cre control and PCC mice (n=4 Cas9 eGFP ; Prx1 -cre and n=6 PCC). (B) Quantification of fibrous layer thickness (μm) at the mid-diaphysis, ROI represented in (A). (C) Representative images for Periostin (POSTN, magenta) and Ki67 (proliferation, green) immunostaining of longitudinal radius sections at two months (n=6 Cas9 eGFP ; Prx1 -cre and n=8 PCC). (D) Quantification of proliferation index (% Ki67+) at the mid-diaphysis, ROIs represented in (C). Significant periosteal expansion results from increased proliferation of cells in the cambium layer. (E) PECAM staining of longitudinal radius sections at two months (n=4 Cas9 eGFP ;Prx1- cre and n=5 PCC). (F) Representative images for RUNX2 (magenta) and Ki67 (green) immunostaining at the mid-diaphysis (n=4 Cas9 eGFP ; Prx1 -cre and n=4 PCC). (G) Quantification of RUNX2+ (magenta bars), Ki67+ (green bars), and RUNX2+Ki67+ double positive cells (magenta and green striped bars) in ROIs represented in (F). (H) Representative images for <t>COL1A1</t> (magenta) and Ki67 (green) immunostaining at the mid-diaphysis (n=6 Cas9 eGFP ; Prx1 -cre and n=6 PCC). (I) Quantification of COL1A1+ (magenta bars), Ki67+ (green bars), and Col1a1+Ki67+ double positive cells (magenta and green striped bars) in ROIs represented in (H). B = bone. c = cambium layer. f = fibrous layer. Ma = marrow. Scale bars = 50 μm. Arrowheads indicate Ki67+ cells. Data represent mean +/-SD. * denotes Welch’s t-test <0.05. ** denotes Welch’s t-test <0.005. *** denotes Welch’s t-test <0.0005. ns = not significant.
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(A) Representative images of SCA-1 immunostaining of longitudinal radius sections at two months focused on the mid-diaphysis of Cas9 eGFP ; Prx1 -cre control and PCC mice (n=4 Cas9 eGFP ; Prx1 -cre and n=6 PCC). (B) Quantification of fibrous layer thickness (μm) at the mid-diaphysis, ROI represented in (A). (C) Representative images for Periostin (POSTN, magenta) and Ki67 (proliferation, green) immunostaining of longitudinal radius sections at two months (n=6 Cas9 eGFP ; Prx1 -cre and n=8 PCC). (D) Quantification of proliferation index (% Ki67+) at the mid-diaphysis, ROIs represented in (C). Significant periosteal expansion results from increased proliferation of cells in the cambium layer. (E) PECAM staining of longitudinal radius sections at two months (n=4 Cas9 eGFP ;Prx1- cre and n=5 PCC). (F) Representative images for RUNX2 (magenta) and Ki67 (green) immunostaining at the mid-diaphysis (n=4 Cas9 eGFP ; Prx1 -cre and n=4 PCC). (G) Quantification of RUNX2+ (magenta bars), Ki67+ (green bars), and RUNX2+Ki67+ double positive cells (magenta and green striped bars) in ROIs represented in (F). (H) Representative images for <t>COL1A1</t> (magenta) and Ki67 (green) immunostaining at the mid-diaphysis (n=6 Cas9 eGFP ; Prx1 -cre and n=6 PCC). (I) Quantification of COL1A1+ (magenta bars), Ki67+ (green bars), and Col1a1+Ki67+ double positive cells (magenta and green striped bars) in ROIs represented in (H). B = bone. c = cambium layer. f = fibrous layer. Ma = marrow. Scale bars = 50 μm. Arrowheads indicate Ki67+ cells. Data represent mean +/-SD. * denotes Welch’s t-test <0.05. ** denotes Welch’s t-test <0.005. *** denotes Welch’s t-test <0.0005. ns = not significant.
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Image Search Results


(A) Representative images of SCA-1 immunostaining of longitudinal radius sections at two months focused on the mid-diaphysis of Cas9 eGFP ; Prx1 -cre control and PCC mice (n=4 Cas9 eGFP ; Prx1 -cre and n=6 PCC). (B) Quantification of fibrous layer thickness (μm) at the mid-diaphysis, ROI represented in (A). (C) Representative images for Periostin (POSTN, magenta) and Ki67 (proliferation, green) immunostaining of longitudinal radius sections at two months (n=6 Cas9 eGFP ; Prx1 -cre and n=8 PCC). (D) Quantification of proliferation index (% Ki67+) at the mid-diaphysis, ROIs represented in (C). Significant periosteal expansion results from increased proliferation of cells in the cambium layer. (E) PECAM staining of longitudinal radius sections at two months (n=4 Cas9 eGFP ;Prx1- cre and n=5 PCC). (F) Representative images for RUNX2 (magenta) and Ki67 (green) immunostaining at the mid-diaphysis (n=4 Cas9 eGFP ; Prx1 -cre and n=4 PCC). (G) Quantification of RUNX2+ (magenta bars), Ki67+ (green bars), and RUNX2+Ki67+ double positive cells (magenta and green striped bars) in ROIs represented in (F). (H) Representative images for COL1A1 (magenta) and Ki67 (green) immunostaining at the mid-diaphysis (n=6 Cas9 eGFP ; Prx1 -cre and n=6 PCC). (I) Quantification of COL1A1+ (magenta bars), Ki67+ (green bars), and Col1a1+Ki67+ double positive cells (magenta and green striped bars) in ROIs represented in (H). B = bone. c = cambium layer. f = fibrous layer. Ma = marrow. Scale bars = 50 μm. Arrowheads indicate Ki67+ cells. Data represent mean +/-SD. * denotes Welch’s t-test <0.05. ** denotes Welch’s t-test <0.005. *** denotes Welch’s t-test <0.0005. ns = not significant.

Journal: bioRxiv

Article Title: CRISPR-mediated conditional mutagenesis of Smad1/5/8 reveals BMP/GDF signaling restricts postnatal bone overgrowth

doi: 10.64898/2026.01.22.701170

Figure Lengend Snippet: (A) Representative images of SCA-1 immunostaining of longitudinal radius sections at two months focused on the mid-diaphysis of Cas9 eGFP ; Prx1 -cre control and PCC mice (n=4 Cas9 eGFP ; Prx1 -cre and n=6 PCC). (B) Quantification of fibrous layer thickness (μm) at the mid-diaphysis, ROI represented in (A). (C) Representative images for Periostin (POSTN, magenta) and Ki67 (proliferation, green) immunostaining of longitudinal radius sections at two months (n=6 Cas9 eGFP ; Prx1 -cre and n=8 PCC). (D) Quantification of proliferation index (% Ki67+) at the mid-diaphysis, ROIs represented in (C). Significant periosteal expansion results from increased proliferation of cells in the cambium layer. (E) PECAM staining of longitudinal radius sections at two months (n=4 Cas9 eGFP ;Prx1- cre and n=5 PCC). (F) Representative images for RUNX2 (magenta) and Ki67 (green) immunostaining at the mid-diaphysis (n=4 Cas9 eGFP ; Prx1 -cre and n=4 PCC). (G) Quantification of RUNX2+ (magenta bars), Ki67+ (green bars), and RUNX2+Ki67+ double positive cells (magenta and green striped bars) in ROIs represented in (F). (H) Representative images for COL1A1 (magenta) and Ki67 (green) immunostaining at the mid-diaphysis (n=6 Cas9 eGFP ; Prx1 -cre and n=6 PCC). (I) Quantification of COL1A1+ (magenta bars), Ki67+ (green bars), and Col1a1+Ki67+ double positive cells (magenta and green striped bars) in ROIs represented in (H). B = bone. c = cambium layer. f = fibrous layer. Ma = marrow. Scale bars = 50 μm. Arrowheads indicate Ki67+ cells. Data represent mean +/-SD. * denotes Welch’s t-test <0.05. ** denotes Welch’s t-test <0.005. *** denotes Welch’s t-test <0.0005. ns = not significant.

Article Snippet: The following primary antibodies and dilutions were used for immunofluorescence of tissue sections: SCA-1(Invitrogen, 14-5981-82, 1: 250); POSTN (Abcam, ab14041, 1: 100) [citric acid]; PECAM (Invitrogen, 14-0311-82, 1: 250); GFP (Invitrogen, A10262, 1: 500); RUNX22 (Abcam, ab76956, 1:500); COL1A1 (Developmental Studies Hybridoma Bank, Sp1.D8, 1:20) [citric acid]; Ki67 rat (Invitrogen, 14-5698-82, 1: 150); Ki67 rabbit (Abcam, ab15580, 1:200); PRX1 (Invitrogen, PA5-106700, 1: 100).

Techniques: Immunostaining, Control, Staining