Journal: BMC Complementary Medicine and Therapies

Article Title: Polygonum multiflorum extract support hair growth by elongating anagen phase and abrogating the effect of androgen in cultured human dermal papilla cells

doi: 10.1186/s12906-020-02940-5

Figure Lengend Snippet: Effect of PM extract on DPCs’ protein expression related to hair cycle. DPCs (1 × 10 6 cells/100 mm dish) were treated with 1 μg/ml, 10 μg/ml and 100 μg/ml of PM extract for 24 h. For DP spheroids, 10 μg/ml and 20 μg/ml of PM extract were treated at the start of spheroid formation. a The protein level of DKK-1 in whole cell lysate of 2D cultured human DPCs and ( b ) supernatant. c Relative mRNA expression of Bcl2 and BAD in 2D cultured DPCs. d Bcl2 expression in 3D DP spheroids. e Bcl2 expression in cryo-sectioned 3D DP spheroids. Data are presented as mean ± SD. n > 3 for each group. * p < 0.05, ** p < 0.01 compared with control group

Article Snippet: TaqMan probes used in this study were as follows: GAPDH assay id 4352934E; BAD assay id Hs00188930_m1; Bcl2 assay id Hs00608023_m1.

Techniques: Expressing, Cell Culture, Control

Journal: Molecular pharmaceutics

Article Title: p53-Bad* Fusion Gene Therapy Induces Apoptosis In Vitro and Reduces Zebrafish Tumor Burden in Hepatocellular Carcinoma

doi: 10.1021/acs.molpharmaceut.2c00665

Figure Lengend Snippet: Characterization of p53-Bad* effects on HCC tumors in FIB p53-Bad* zebrafish. (A) p53-Bad* effects on HCC tumor burden as measured by the liver–body mass ratio. N = 40 (WT), N = 51 (p53-Bad*), N = 40 (pt-β-cat), and N = 46 (pt-β-cat/p53-Bad*). One-way ANOVA (p < 0.0001) with Tukey’s post-test was performed: ns = not significant, ****p < 0.0001. (B) p53-Bad* effects on liver histology for p53-Bad* Founder B. N = 40 (WT), N = 51 (p53-Bad*), N = 40 (pt-β-cat), and N = 46 (pt-β-cat/p53-Bad*). Fisher’s exact test between pt-β-cat and pt-β-cat/p53-Bad* groups for HCC positive versus HCC negative histology gave a p-value = 0.04. (C) Examples of male zebrafish with no significant pathological abnormality (top panel)—nuclei are uniform, round, and regularly spaced, and architecture is normal, including scattered bile ducts (inset); mild changes (middle panel)—architecture is normal, including scattered bile ducts (arrow), but with occasional enlarged nuclei (inset); and HCC (bottom panel)—the architecture is disorganized, and nuclei are enlarged, pleomorphic, and irregular (inset).

Article Snippet: Pre-designed Taqman probes (ThermoFisher) targeted human 18S (Hs99999901_sl), human Bad (Hs00997773_ml), human p53 (Hs01034254_gl TP53), human Mcl-1 (Hs01050896_ml MCLl), human Bcl-2 (Hs01048932_gl BCL2), human Bcl-xL (Hs00236329_ml BCL2L1), and zebrafish beta-actin (Dr03432610_ml actbl).

Techniques:

Journal: Molecular pharmaceutics

Article Title: p53-Bad* Fusion Gene Therapy Induces Apoptosis In Vitro and Reduces Zebrafish Tumor Burden in Hepatocellular Carcinoma

doi: 10.1021/acs.molpharmaceut.2c00665

Figure Lengend Snippet: Microscopy of HCC cell lines to determine mitochondrial localization. (A) Representative cells from C3A (WT p53), Hep3B2.1–7 (p53 null), and PLC/PRF/5 (p53 R249S) transfected with CMV-GFP, CMV-GFP-Bad, CMV-GFP-Bad* CMV-GFP-p53, CMV-GFP-p53-Bad, and CMV-GFP-p53-Bad*. The nucleus is visualized in blue (Hoechst), the mitochondria in red (MitoTracker), and each construct in green (GFP). (B) PCC colocalization analyses for red (mitochondria) vs green (construct). N ≥ 30 cells for each construct with post-Costes’ automatic threshold algorithm, PCC > 0.6 considered colocalized. One-way ANOVA with Tukey’s post-test was performed for each graph: ns = not significant, **p < 0.01, and ****p < 0.0001.

Article Snippet: Pre-designed Taqman probes (ThermoFisher) targeted human 18S (Hs99999901_sl), human Bad (Hs00997773_ml), human p53 (Hs01034254_gl TP53), human Mcl-1 (Hs01050896_ml MCLl), human Bcl-2 (Hs01048932_gl BCL2), human Bcl-xL (Hs00236329_ml BCL2L1), and zebrafish beta-actin (Dr03432610_ml actbl).

Techniques: Microscopy, Transfection, Construct

Journal: Molecular pharmaceutics

Article Title: p53-Bad* Fusion Gene Therapy Induces Apoptosis In Vitro and Reduces Zebrafish Tumor Burden in Hepatocellular Carcinoma

doi: 10.1021/acs.molpharmaceut.2c00665

Figure Lengend Snippet: Apoptosis in p53-Bad*/pt-β-cat zebrafish. (A) Detection of apoptotic bodies via the TUNEL assay in zebrafish liver cross-sections. N = 5 (HCC) and N = 6 (other groups). One-way ANOVA (p = 0.0021) with Tukey’s post-test was performed: ns = not significant, *p < 0.05, **p < 0.01. (B) Representative images of TUNEL staining; arrows point to apoptotic bodies.

Article Snippet: Pre-designed Taqman probes (ThermoFisher) targeted human 18S (Hs99999901_sl), human Bad (Hs00997773_ml), human p53 (Hs01034254_gl TP53), human Mcl-1 (Hs01050896_ml MCLl), human Bcl-2 (Hs01048932_gl BCL2), human Bcl-xL (Hs00236329_ml BCL2L1), and zebrafish beta-actin (Dr03432610_ml actbl).

Techniques: TUNEL Assay, Staining

Journal: Molecular pharmaceutics

Article Title: p53-Bad* Fusion Gene Therapy Induces Apoptosis In Vitro and Reduces Zebrafish Tumor Burden in Hepatocellular Carcinoma

doi: 10.1021/acs.molpharmaceut.2c00665

Figure Lengend Snippet: p53-Bad* and mitochondrial apoptosis. (A) When active, pro-apoptotic proteins Bak and Bax homo-oligomerize and induce formation of mitochondrial pores, releasing cytochrome c. This leads to caspase cascade activation, apoptosome formation, and induction of apoptosis. (B) Anti-apoptotic factors such as Bcl-xL, Bcl-2, and Mcl-1 bind to Bak and/or Bax, preventing homo-oligomerization. At the mitochondria, p53 (C) inactivates anti-apoptotic factors and (D) activates Bak and Bax. (E) Pro-apoptotic BH3-only proteins such as Bad bind to anti-apoptotic factors, freeing Bak and Bax. (F) p53-Bad* localizes to the mitochondria, enabling the dual activity of p53 and Bad.

Article Snippet: Pre-designed Taqman probes (ThermoFisher) targeted human 18S (Hs99999901_sl), human Bad (Hs00997773_ml), human p53 (Hs01034254_gl TP53), human Mcl-1 (Hs01050896_ml MCLl), human Bcl-2 (Hs01048932_gl BCL2), human Bcl-xL (Hs00236329_ml BCL2L1), and zebrafish beta-actin (Dr03432610_ml actbl).

Techniques: Activation Assay, Activity Assay

Journal: Redox Report : Communications in Free Radical Research

Article Title: Sicyos angulatus ameliorates acute liver injury by inhibiting oxidative stress via upregulation of anti-oxidant enzymes

doi: 10.1080/13510002.2018.1546986

Figure Lengend Snippet: Effect of SA on histological alterations and apoptosis in the liver after Con A exposure. Histological assessments were performed by H&E (magnification: 200× or 400×) (A) and TUNEL staining to detect apoptotic cells (magnification: 100×) (B). Scale bars, 100 μm or 200 μm. * p < 0.05 compared to Con A + Veh mice by Student’s t -test. Injury area and necrotic area presented as the percentage of liver section.

Article Snippet: Concentration-dependent induction of (A) markers of apoptosis (Bax, Bcl-2, Bad, and Bim) in the HepG2 cells following treatment with SA and (B) mitochondrial release of apoptosis-related proteins (Smac/DIABLO, cytochrome C, and AIF) were detected in the cytosolic cell fractions by western blotting. β-actin was used as a loading control for each experiment.

Techniques: TUNEL Assay, Staining

Journal: Redox Report : Communications in Free Radical Research

Article Title: Sicyos angulatus ameliorates acute liver injury by inhibiting oxidative stress via upregulation of anti-oxidant enzymes

doi: 10.1080/13510002.2018.1546986

Figure Lengend Snippet: Effects of SA on apoptotic protein levels and neutrophil infiltration in Con A-treated HepG2 cells. The expression levels of apoptotic proteins was measured by western blotting. Concentration-dependent induction of (A) markers of apoptosis (Bax, Bcl-2, Bad, and Bim) in the HepG2 cells following treatment with SA and (B) mitochondrial release of apoptosis-related proteins (Smac/DIABLO, cytochrome C, and AIF) were detected in the cytosolic cell fractions by western blotting. β-actin was used as a loading control for each experiment. The data are representatives of three independent experiments. (C) Histological assessment was performed by immunohistochemical staining using an antibody against neutrophils, and representative images are displayed (original magnification: 100×). Arrows indicate neutrophils. Neutrophil counts in the liver tissues (right grape). ** Significantly different from the group treated with Con A-Veh, p < 0.01 (Student’s t -test).

Article Snippet: Concentration-dependent induction of (A) markers of apoptosis (Bax, Bcl-2, Bad, and Bim) in the HepG2 cells following treatment with SA and (B) mitochondrial release of apoptosis-related proteins (Smac/DIABLO, cytochrome C, and AIF) were detected in the cytosolic cell fractions by western blotting. β-actin was used as a loading control for each experiment.

Techniques: Expressing, Western Blot, Concentration Assay, Control, Immunohistochemical staining, Staining