Journal: Cells

Article Title: Regulation of Neurogenesis in Mouse Brain by HMGB1

doi: 10.3390/cells9071714

Figure Lengend Snippet: ( A ) Sagittal sections of E16 embryos with BrdU staining. Scale bar indicates 500 µm. The black rectangles are the areas shown in B. ( B ) BrdU staining of E16 dorsal telencephalon under higher resolution. Scale bar indicates 100 µm. ( C ) Statistics of the BrdU + cell numbers in the E16 cortical layers of the KO and WT embryos. For each brain, BrdU + cell numbers in each layer were counted five times at different sections for averaging. The stained sections from 5 KO and 5 WT embryos were used for comparison. Mean values ± S.D. (error bars) and S.E.M (Boxes) are indicated ( n = 5; *** p < 0.001; * p < 0.05, unpaired t -test, two-tailed). ( D ) Sagittal sections of E16 embryos with hematoxylin staining. The areas in red rectangle were zoomed out in ( E ) Scale bar indicates 500 µm. ( E ) The E16 cortical layers in the dorsal pallium (Dpa) with hematoxylin stains. Scale bar indicates 150 µm. ( F ) Statistics of the thickness of E16 cortical layers. Each layer has been measured on three different sections at central position for each embryo. Sections from six KO and six WT mice have been measured. Mean values ± S.D. (error bars) and S.E.M (Boxes) are indicated ( n = 6; **, p < 0.01; *, p < 0.05, unpaired t -test, two-tailed). ( G ) The Tunel staining of E12 sagittal sections. The strong apoptotic signaling in the dorsal neopallial cortex (NPC) and subpallium (Spa) of the HMGB1 KO is indicated by white arrows. The area in the white rectangle has been amplified. The intermediate layer of NPC showed highest apoptotic activity (***). Scale bar indicates 200 µm in the upper panel and 100 µm in the lower panel. Abbreviations: AOV, ventral anterior olfactory area; CP, cortical plate; Dpa, dorsal pallium/isocortex; H, hindbrain; HI, hippocampus; IZ, intermediate zone; M, midbrain; MAR, marginal layer; NPC, neopallial cortex; Spa, subpallium; STR, striatum; SVZ, subventricular zone; Tel, telencephalic vesicle; VD, ventricles of diencephalon; VZ, ventricular zone.

Article Snippet: The fluorometric TUNEL system (Alexa ® 488, DeadEndTM, Promega) was used for staining of apoptosis in whole mounts.

Techniques: BrdU Staining, Staining, Comparison, Two Tailed Test, TUNEL Assay, Amplification, Activity Assay

Journal: Cells

Article Title: Regulation of Neurogenesis in Mouse Brain by HMGB1

doi: 10.3390/cells9071714

Figure Lengend Snippet: Apoptotic and proliferating activity of in vitro cultured E16 cortical neuronal cells. ( A ) Tunel staining of E16 cortical neuronal cells cultured for 2 days. All of the samples have been co-stained with anti-HMGB1 antibody (Alexa 568) and DAPI. Scale bar indicates 40 μm. ( B ) Tunel staining and anti-PCNA antibody (Alexa 568) staining of E16 cortical neuronal cells cultured for two, four, and seven days. Cell nuclei have been stained with DAPI. Recombinant HMGB1 protein (recHMGB1) was coated on the matrix and added into the culture medium at 10 μg/mL. Scale bar indicates 50 μm. ( A ) and ( B ) are the confocal z-stack max projections of 10 μm. ( C ) The statistics of apoptotic cells in each group of cultured cells shown in ( B ). ( D ) The statistics of proliferating cells in each group of cultured cells shown in ( B ). For the statistics in ( C ) and ( D ), the cortical neurons from six KO and six WT E16 brains were collected and cultured for the immunostaining and analysis. For each brain sample, three repeats were applied. Mean values ± S.D. (error bars) and S.E.M (Boxes) are indicated ( n = 6; *** p < 0.001; ** p < 0.01; * p < 0.05; unpaired t -test, two-tailed).

Article Snippet: The fluorometric TUNEL system (Alexa ® 488, DeadEndTM, Promega) was used for staining of apoptosis in whole mounts.

Techniques: Activity Assay, In Vitro, Cell Culture, TUNEL Assay, Staining, Recombinant, Immunostaining, Two Tailed Test