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a) Intracranial injection of DF1 producing cells carrying RCAS vectors encoding PDGFB, Cre, and luciferase to enable conditional targeting of neural stem cells. b) Mice were injected at postnatal day 3-5 and monitored for tumor development through symptom assessment and bioluminescence imaging starting at week 4 to detect tumor-associated signals. Kaplan-Meier survival estimates and and log-rank (Mantel-Cox) test p= 0.0277, Ppm1d flex- /wt ; p53 fl/wt n=21, Ppm1d wt/wt ; p53 fl/wt n=19 c) Immunohistochemistry <t>(IHC)</t> with <t>Ki67,</t> Olig2, and GFAP for tumor-bearing brains from wild-type Ppm1d (Ppm1d wt/wt ; p53 fl/wt ) and truncated Ppm1d (Ppm1d flex- /wt ; p53 fl/wt ) mice. Tumor and normal regions were denoted in GFAP staining; GFAP exhibited cytoplasmic localization. Scale bar: 100 μm. d) Quantification of Ki67 and Olig2 staining. GFAP expression was excluded from quantification due to its cytoplasmic distribution. Data are presented as mean ± SEM; n = 3. Student paired t-test, * p< 0.05 and ** p< 0.01 e) IHC staining for Myc-tag was performed on brain sections from wild-type Ppm1d (Ppm1d wt/wt ; p53 fl/wt ) and truncated Ppm1d (Ppm1d flex- /wt ; p53 fl/wt ) mice. Whole-mount sections (scale bar 2 mm) and tumor-region sections (scale bar 100 μm). Red boxes represent tumor regions. f) Quantification of Myc Tag IHC staining. Data are shown as mean ± SEM; n = 3. Student unpaired t-test, ** p< 0.01
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a) Intracranial injection of DF1 producing cells carrying RCAS vectors encoding PDGFB, Cre, and luciferase to enable conditional targeting of neural stem cells. b) Mice were injected at postnatal day 3-5 and monitored for tumor development through symptom assessment and bioluminescence imaging starting at week 4 to detect tumor-associated signals. Kaplan-Meier survival estimates and and log-rank (Mantel-Cox) test p= 0.0277, Ppm1d flex- /wt ; p53 fl/wt n=21, Ppm1d wt/wt ; p53 fl/wt n=19 c) Immunohistochemistry <t>(IHC)</t> with <t>Ki67,</t> Olig2, and GFAP for tumor-bearing brains from wild-type Ppm1d (Ppm1d wt/wt ; p53 fl/wt ) and truncated Ppm1d (Ppm1d flex- /wt ; p53 fl/wt ) mice. Tumor and normal regions were denoted in GFAP staining; GFAP exhibited cytoplasmic localization. Scale bar: 100 μm. d) Quantification of Ki67 and Olig2 staining. GFAP expression was excluded from quantification due to its cytoplasmic distribution. Data are presented as mean ± SEM; n = 3. Student paired t-test, * p< 0.05 and ** p< 0.01 e) IHC staining for Myc-tag was performed on brain sections from wild-type Ppm1d (Ppm1d wt/wt ; p53 fl/wt ) and truncated Ppm1d (Ppm1d flex- /wt ; p53 fl/wt ) mice. Whole-mount sections (scale bar 2 mm) and tumor-region sections (scale bar 100 μm). Red boxes represent tumor regions. f) Quantification of Myc Tag IHC staining. Data are shown as mean ± SEM; n = 3. Student unpaired t-test, ** p< 0.01
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a) Intracranial injection of DF1 producing cells carrying RCAS vectors encoding PDGFB, Cre, and luciferase to enable conditional targeting of neural stem cells. b) Mice were injected at postnatal day 3-5 and monitored for tumor development through symptom assessment and bioluminescence imaging starting at week 4 to detect tumor-associated signals. Kaplan-Meier survival estimates and and log-rank (Mantel-Cox) test p= 0.0277, Ppm1d flex- /wt ; p53 fl/wt n=21, Ppm1d wt/wt ; p53 fl/wt n=19 c) Immunohistochemistry <t>(IHC)</t> with <t>Ki67,</t> Olig2, and GFAP for tumor-bearing brains from wild-type Ppm1d (Ppm1d wt/wt ; p53 fl/wt ) and truncated Ppm1d (Ppm1d flex- /wt ; p53 fl/wt ) mice. Tumor and normal regions were denoted in GFAP staining; GFAP exhibited cytoplasmic localization. Scale bar: 100 μm. d) Quantification of Ki67 and Olig2 staining. GFAP expression was excluded from quantification due to its cytoplasmic distribution. Data are presented as mean ± SEM; n = 3. Student paired t-test, * p< 0.05 and ** p< 0.01 e) IHC staining for Myc-tag was performed on brain sections from wild-type Ppm1d (Ppm1d wt/wt ; p53 fl/wt ) and truncated Ppm1d (Ppm1d flex- /wt ; p53 fl/wt ) mice. Whole-mount sections (scale bar 2 mm) and tumor-region sections (scale bar 100 μm). Red boxes represent tumor regions. f) Quantification of Myc Tag IHC staining. Data are shown as mean ± SEM; n = 3. Student unpaired t-test, ** p< 0.01
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a) Intracranial injection of DF1 producing cells carrying RCAS vectors encoding PDGFB, Cre, and luciferase to enable conditional targeting of neural stem cells. b) Mice were injected at postnatal day 3-5 and monitored for tumor development through symptom assessment and bioluminescence imaging starting at week 4 to detect tumor-associated signals. Kaplan-Meier survival estimates and and log-rank (Mantel-Cox) test p= 0.0277, Ppm1d flex- /wt ; p53 fl/wt n=21, Ppm1d wt/wt ; p53 fl/wt n=19 c) Immunohistochemistry (IHC) with Ki67, Olig2, and GFAP for tumor-bearing brains from wild-type Ppm1d (Ppm1d wt/wt ; p53 fl/wt ) and truncated Ppm1d (Ppm1d flex- /wt ; p53 fl/wt ) mice. Tumor and normal regions were denoted in GFAP staining; GFAP exhibited cytoplasmic localization. Scale bar: 100 μm. d) Quantification of Ki67 and Olig2 staining. GFAP expression was excluded from quantification due to its cytoplasmic distribution. Data are presented as mean ± SEM; n = 3. Student paired t-test, * p< 0.05 and ** p< 0.01 e) IHC staining for Myc-tag was performed on brain sections from wild-type Ppm1d (Ppm1d wt/wt ; p53 fl/wt ) and truncated Ppm1d (Ppm1d flex- /wt ; p53 fl/wt ) mice. Whole-mount sections (scale bar 2 mm) and tumor-region sections (scale bar 100 μm). Red boxes represent tumor regions. f) Quantification of Myc Tag IHC staining. Data are shown as mean ± SEM; n = 3. Student unpaired t-test, ** p< 0.01

Journal: bioRxiv

Article Title: Oncogenic Ppm1d mutations deregulate the p53 pathway in primary mouse gliomas

doi: 10.1101/2025.11.25.690483

Figure Lengend Snippet: a) Intracranial injection of DF1 producing cells carrying RCAS vectors encoding PDGFB, Cre, and luciferase to enable conditional targeting of neural stem cells. b) Mice were injected at postnatal day 3-5 and monitored for tumor development through symptom assessment and bioluminescence imaging starting at week 4 to detect tumor-associated signals. Kaplan-Meier survival estimates and and log-rank (Mantel-Cox) test p= 0.0277, Ppm1d flex- /wt ; p53 fl/wt n=21, Ppm1d wt/wt ; p53 fl/wt n=19 c) Immunohistochemistry (IHC) with Ki67, Olig2, and GFAP for tumor-bearing brains from wild-type Ppm1d (Ppm1d wt/wt ; p53 fl/wt ) and truncated Ppm1d (Ppm1d flex- /wt ; p53 fl/wt ) mice. Tumor and normal regions were denoted in GFAP staining; GFAP exhibited cytoplasmic localization. Scale bar: 100 μm. d) Quantification of Ki67 and Olig2 staining. GFAP expression was excluded from quantification due to its cytoplasmic distribution. Data are presented as mean ± SEM; n = 3. Student paired t-test, * p< 0.05 and ** p< 0.01 e) IHC staining for Myc-tag was performed on brain sections from wild-type Ppm1d (Ppm1d wt/wt ; p53 fl/wt ) and truncated Ppm1d (Ppm1d flex- /wt ; p53 fl/wt ) mice. Whole-mount sections (scale bar 2 mm) and tumor-region sections (scale bar 100 μm). Red boxes represent tumor regions. f) Quantification of Myc Tag IHC staining. Data are shown as mean ± SEM; n = 3. Student unpaired t-test, ** p< 0.01

Article Snippet: IHC staining for Ki67 (Cat #Bethyl IHC-00375), Olig2 (Cat #ab109186), GFAP (Cat #NB300-141) and MYC (Cat #ab32072) was conducted by HistoWiz.

Techniques: Injection, Luciferase, Imaging, Immunohistochemistry, Staining, Expressing