p16 3mr transgenic mice  (Jackson Laboratory)

Journal: bioRxiv

Article Title: Gut Microbiota-derived Acetate Safeguards the Colonic Epithelial Acetyl-CoA Reserve to Avert Colonic Senescence

doi: 10.64898/2026.05.15.725523

Figure Lengend Snippet: (A) Schematic illustration of the tri-modality reporter in p16-3MR transgenic mice and the generation of CEC-Acly −/− ;p16-3MR mice; (B) SA-β-gal staining in frozen colon sections from Acly f/f ;p16-3MR and CEC- Acly −/− ;p16-3MR mice on days 0, 7 and 10 post ABX; (C) Luciferase (Luc) activity in purified colonic crypts from Acly f/f ;p16-3MR and CEC- Acly −/− ;p16-3MR mice on days 0, 7 and 10 post ABX; (D) RFP-positive cells in colonic mucosa from Acly f/f ;p16-3MR and CEC- Acly −/− ;p16-3MR mice on days 0, 7 and 10 post ABX; (E) Kaplan-Meier survival curves of Acly f/f ;p16-3MR and CEC- Acly −/− ;p16-3MR mice following ABX and ganciclovir (GCV) treatment; (F) Alcian blue stained colon sections of Acly f/f ;p16-3MR and CEC- Acly −/− ;p16-3MR mice on day 10 after ABX and PBS treatment, and on day 18 after ABX and GCV treatment; (G) Luciferase (Luc) activity in purified colonic crypt lysates from Acly f/f ;p16-3MR and CEC- Acly −/− ;p16-3MR mice on day 10 post ABX and PBS or GCV treatment; (H) RFP-positive cells in colonic mucosa from Acly f/f ;p16-3MR and CEC- Acly −/− ;p16-3MR mice on day 10 post ABX and PBS or GCV treatment; (I) Serum cytokine concentrations in Acly f/f ;p16-3MR and CEC- Acly −/− ;p16-3MR mice on day 10 post ABX and PBS or GCV treatment; (J) Western blot assessment of senescence hallmarks in colonic crypt cells from Acly f/f ;p16-3MR and CEC- Acly −/− ;p16-3MR mice on day 10 post ABX and PBS or GCV treatment. Data are presented as mean ± SD. *p<0.05; **p<0.01; ***p<0.001, ****p<0.0001 by two-way ANOVA.

Article Snippet: CDX2-Cre transgenic mice (B6.Cg-Tg(CDX2-cre)101Erf/J; Stock # 009350), CDX2-CreERT2 transgenic mice (B6.Cg-Tg(CDX2-Cre/ERT2)752Erf/J; Stock # 022390), p16-3MR transgenic mice (B6.Cg-Tg(Cdkn2a/luc/RFP/TK)1Cmps/J, Stock # 037045) and Trp53 −/− mice (B6.129S2- Trp53 tm1Tyj /J; Stock # 002101) were purchased from Jackson Laboratory.

Techniques: Transgenic Assay, Staining, Luciferase, Activity Assay, Purification, Western Blot

Journal: Nature medicine

Article Title: Local clearance of senescent cells attenuates the development of post-traumatic osteoarthritis and creates a pro-regenerative environment

doi: 10.1038/nm.4324

Figure Lengend Snippet: Clearance of SnCs by GCV reduces the development of post-traumatic OA. (a) Schematic of the time course for the experiments in b–h. Male p16-3MR mice undergoing ACLT were injected intra-articularly with vehicle (Veh) or ganciclovir (GCV) and evaluated as indicated. (b) Representative luminescence images (sham surgery, n = 7; Veh treated, n = 7; GCV treated, n = 3) of ACLT mice after vehicle or GCV treatment on day 28 after surgery (left) and quantification of the luminescence (right) at the indicated time s(in arbitrary units, A.U.). Scale bars, 2 cm. (c) Quantification of mRNA expression for Cdkn2a in articular joints treated on day 28 after surgery (no surgery, sham surgery and Veh treated, n = 3; GCV treated, n = 5). (d) Quantification of mRNA expression for Cdkn1a, Il1b, Mmp13, Col2a1, Acan and Sox9, normalized to Actb expression, in joints from no-surgery, sham-operated and ACLT mice that were treated as indicated (no surgery, sham surgery and Veh treated, n = 3; GCV treated, n = 5). (e) Representative images of HMGB1 (brown, closed arrowheads; no surgery, sham surgery and GCV treated, n = 3; Veh treated, n = 4) and p16INK4a (brown, open arrowheads; no surgery and sham surgery, n = 3; Veh treated, n = 5; GCV treated, n = 4) immunostaining and safranin O and methyl green (no surgery, n = 3; sham surgery, n = 5; Veh treated, n = 6; GCV treated, n = 8) in p16-3MR mice. F, femur; T, tibia; M, meniscus. Scale bars, 100 μm. (f) Quantification of non-SnCs positive for nuclear HMGB1 (no surgery and sham surgery, n = 3; Veh treated, n = 4; GCV treated, n = 3) and SnCs positive for p16INK4a (no surgery and sham surgery, n = 3; Veh treated, n = 5; GCV treated, n = 4) in articular cartilage. (g) Medial tibial plateau joint score in p16-3MR mice based on the OARSI scoring system (no surgery, n = 3; sham surgery, n = 5; Veh treated, n = 6; GCV treated, n = 8). (h) The percentage of weight placed on the operated limb versus the contralateral control (top) and the response time of mice after placement onto a 55 °C hotplate (bottom) (no surgery, sham surgery and Veh treated, n = 5; GCV treated, n = 10). All data are expressed as means, and each data point represents an individual mouse. One-way ANOVA with Tukey’s multiple-comparisons test was used for statistical analysis in c and d; a two-tailed t-test (unpaired) was used for b and f–h. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; N.S., not significant.

Article Snippet: Mice generation and drug treatment We purchased young (aged 10 weeks) male C57BL mice from Charles River (Germantown, MD, USA). p16-3MR transgenic mice (a C57BL/6J strain) were generated as previously described 15 and were bred in the animal facility of the Johns Hopkins University School of Medicine.

Techniques: Injection, Expressing, Immunostaining, Control, Two Tailed Test

Journal: Nature medicine

Article Title: Local clearance of senescent cells attenuates the development of post-traumatic osteoarthritis and creates a pro-regenerative environment

doi: 10.1038/nm.4324

Figure Lengend Snippet: Clearance of SnCs slows the development of naturally occurring OA and post-traumatic OA in aged mice. (a) Study design for clearance of naturally occurring p16INK4a-positive SnCs in female INK-ATTAC mice. AP20187 (AP) was administrated intraperitoneally (i.p.) twice a week starting at 12 months of age. (b) Representative images of safranin O and methyl green staining from vehicle-treated (−AP; n = 6) and AP20187-treated (+AP; n = 4) mice. F, femur; T, tibia. Scale bars, 100 μm. (c) OARSI grade for vehicle-treated (n = 6 mice aged 20, 21, 25, 28, 28 and 32 months) and AP20187-treated (n = 4 mice aged 21, 28, 28 and 34 months) mice. (d) Representative whole-body luminescence images on day 28 after ACLT surgery (no surgery, n = 8; Veh treated, n = 7; UBX0101 treated, n = 8) (left) and quantification of luminescence (in arbitrary units, A.U.) at the indicated times after the surgery of male p16-3MR mice aged 19–20 months treated as indicated in Figure 2a (right). Scale bars, 2 cm (no surgery, n = 8; Veh treated, n = 7; UBX treated, n = 8). (e) Quantification of mRNA levels for Cdkn2a, Cdkn1a, Mmp13, Col2a1 and Acan normalized to Actb levels in joints 28 d after ACLT (n = 3 for all groups). (f) Representative images of safranin O and methyl green staining and immunostaining for p16INK4a (brown, arrows; no surgery, n = 4; Veh treated, n = 3; UBX0101 treated, n = 4), Ki-67 (no surgery and Veh treated, n = 3; UBX0101 treated, n = 4) and HMGB1 (brown, arrowheads; no surgery, n = 5; Veh treated and UBX0101 treated, n = 3) on articular cartilage. HC, hyaline cartilage; CC, calcified cartilage. Scale bars, 100 μm. (g) OARSI grade for no-surgery, vehicle-treated and UBX0101-treated ACLT mice (no surgery, n = 5; Veh treated, n = 6; UBX treated, n = 7). (h) The percentage of weight placed on the operated limb versus the contralateral control (left) and the response time of mice after placement onto a 55 °C platform (right) (no surgery, n = 6; Veh treated, n = 10; UBX treated, n = 11). All data are expressed as means, and each data point represents an individual mouse. One-way ANOVA with Tukey’s multiple-comparisons test was used for statistical analysis in e; a two-tailed t-test (unpaired) was used for c, d, g and h. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Mice generation and drug treatment We purchased young (aged 10 weeks) male C57BL mice from Charles River (Germantown, MD, USA). p16-3MR transgenic mice (a C57BL/6J strain) were generated as previously described 15 and were bred in the animal facility of the Johns Hopkins University School of Medicine.

Techniques: Staining, Immunostaining, Control, Two Tailed Test