Journal: Physiological reports

Article Title: Hyperoxia causes senescence and increases glycolysis in cultured lung epithelial cells.

doi: 10.14814/phy2.14839

Figure Lengend Snippet: FIGURE 2 Hyperoxia causes DNA damage and p53 activation. Mouse lung epithelial (MLE-12) cells were exposed to 21% O2/5% CO2 (Air) or 95% O2/5% CO2 (O2) for 24 h. For immunofluorescence, Blue = DAPI, nuclei; Green = protein of interest, Cyan = overlapping blue and green signal. (a) Measured γH2AX levels using immunofluorescence. (b) Measured 53BP1 levels using immunofluorescence, quantified by macros script. (c) Transcriptional levels of p53, pRB, p16, and p21 measured by RT-qPCR. (d, e) p53 and pRB protein levels measured by western blot. Western blot normalized using calnexin (CNX) levels. (f) p53 nuclear and cytoplasmic levels measured by western blot from nuclear and cytoplasmic extracts. (g) Nuclear quantification of p53 using immunofluorescence. (h) Levels of phosphorylated p53 at Ser15 as measured by ELISA. Sample turns blue according to phosphorylated levels of p53 at Ser15, absorbance measured at 450 nm. *p < 0.05 versus air groups. RT- qPCR, quantitative real time PCR

Article Snippet: Primary antibodies included p53 (abcam 31333 at 1:200 dilution), phospho- histone H2A Ser139 (Millipore 05- 636 at 1:100 dilution) and 53BP1 (Novus Biologicals NB100- 304 at 1:1000 dilution).

Techniques: Activation Assay, Immunofluorescence, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction

Journal: Physiological reports

Article Title: Hyperoxia causes senescence and increases glycolysis in cultured lung epithelial cells.

doi: 10.14814/phy2.14839

Figure Lengend Snippet: FIGURE 3 Hyperoxia-induced senescence is p53-dependent and pRB-independent. (a) Cas9 knock-in, p53KO, and pRBKO shown by Western blot, and calnexin (CNX) as a housekeeping control. (b) Senescence levels using C12FDG FACS of WT, pRBKO, and p53KO MLE-12 cells exposed to 21% O2/5% CO2 (Air, white) or 95% O2/5% CO2 (O2, black) for 24 h. (c) Emission spectrum of senescent cells is shown in blue for Air, and O2 is in red. (d) γH2AX and (e) 53BP1 levels were measured using immunofluorescence. Blue = DAPI, nuclei; Green = protein of interest, Cyan = overlapping blue and green signal. (f) γH2AX protein levels were measured by Western blot in p53KO and WT cells exposed to hyperoxia. (g) Transcriptional levels of p21, pRB, E2F1, p16, MMP3, MMP10 measured by RT-qPCR. *p < 0.05. MLE-12, mouse lung epithelial

Article Snippet: Primary antibodies included p53 (abcam 31333 at 1:200 dilution), phospho- histone H2A Ser139 (Millipore 05- 636 at 1:100 dilution) and 53BP1 (Novus Biologicals NB100- 304 at 1:1000 dilution).

Techniques: Knock-In, Western Blot, Control, Immunofluorescence, Quantitative RT-PCR

Journal: Frontiers in pharmacology

Article Title: Ginsenoside Rb1 Lessens Gastric Precancerous Lesions by Interfering With β-Catenin/TCF4 Interaction.

doi: 10.3389/fphar.2021.682713

Figure Lengend Snippet: FIGURE 6 | GRb1 inhibits hyper-proliferation of epithelial cells and accelerates apoptosis in GPL model rats. Representative images showing (A) PCNA expression and (C) Ki-67 expression as well as (E) TUNEL apoptotic cells in gastric tissue sections from each group (magnification ×100 and ×400). Semi-quantitative analysis of (B) PCNA protein expression levels and (D) Ki-67 protein expression levels as well as (F) TUNEL apoptosis ratio in each group (n 10). *p < 0.05 and **p < 0.01 vs. Normal group. #p < 0.05 and ##p < 0.01 vs. Model group. Data are presented as mean ± SEM. Abbreviations: GRb1, ginsenoside Rb1; GPL, gastric precancerous lesions; PCNA, proliferating cell nuclear antigen; TUNEL, TdT-mediated dUTP nick-end labeling; SEM, standard error of mean.

Article Snippet: Ki67 antibody was supplied by Novus Biologicals, LLC, United States (cat. no. NB500-170).

Techniques: Expressing, TUNEL Assay, End Labeling