Journal: EBioMedicine

Article Title: Disruption of FDPS/Rac1 axis radiosensitizes pancreatic ductal adenocarcinoma by attenuating DNA damage response and immunosuppressive signalling

doi: 10.1016/j.ebiom.2021.103772

Figure Lengend Snippet: Zol and RT treatment modulate DNA damage response by deactivating Rho small GTPases, thereby reducing DNA double-stranded break repair proteins. (a) Venn diagram showing specific differentially expressed genes in different treatment groups. (b) Pathway enrichment analysis of common and unique genes in the RT versus Zol+RT group. (c) Integrated network analysis of differentially expressed genes in the Zol+RT versus RT group using STRING version 10.5. (d) Dose-dependent treatment of PDAC cells with Zol decreased Rac-1 activity. (e) Western blot analysis confirmed the expression of prenylated (Arrows in red) and unprenylated (Arrows in green) forms of Ras family members Rac-1 and CDC42 upon dose-dependent treatment of Zol in BxPC3, T3M4 and CD18/HPAF PDAC cells. (f) Human and mouse PDAC cells were exposed to Zol (5 µM) 4 h prior to RT, lysates were prepared, and proteins were separated using SDS-PAGE and analysed for antibodies specific for small GTPases (Rac-1, CDC42, and Rho G) and DNA repair and damage signalling (RAD50, MRE11, pBRCA1, and KU80) (g and h) Transient knockdown (KD) of FDPS in PDAC cells suppressed FDPS and subsequently reduced RAD50. (g) Clonogenic survival assay confirmed that FDPS suppression reduced the number of colonies as quantified using Image J software. The bar graph represents the results of the mean ±SE of colonies that survived after FDPS KD and RT (SCR vs. SCR+RT ( P = 0.034), Si-A vs. Si-A+RT ( P < 0.001), Si-B vs. Si-B+RT ( P = 0.036), SCR+RT vs. Si-A+RT ( P = 0.0072), Si-A+RT vs. Si-B+RT ( P = 0.049), SCR+RT vs. Si-B+RT ( P = 0.0048), n=3, student's t-test ). (h) FDPS transient transfection potency was verified using lysates prepared from transfection of non-targeting (Scramble) and siRNA targeting FDPS in PDAC cells and analysed for FDPS, RAD50, and β-actin protein by western blot analysis.

Article Snippet: These membranes were probed with the following primary antibodies with respective dilutions overnight at 4°C: FDPS (mouse, 1:1000, proteintech # 16129-1-AP), pERK (rabbit, 1:1000, Cell signaling technology # 9101, Danvers, MA, USA), ERK (mouse, 1:2000, Cell signaling technology # 9102), anti-β-actin (mouse, 1:5000, Sigma # A1978, St Louis, MO, USA), and GAPDH.

Techniques: Activity Assay, Western Blot, Expressing, SDS Page, Knockdown, Clonogenic Cell Survival Assay, Software, Transfection

Journal: Frontiers in Pharmacology

Article Title: Transcriptomic Analysis Reveals the Protective Effects of Empagliflozin on Lipid Metabolism in Nonalcoholic Fatty Liver Disease

doi: 10.3389/fphar.2021.793586

Figure Lengend Snippet: The sequence of the primers for qRT-PCR.

Article Snippet: After blocking with 5% BSA, the membranes were incubated with primary antibodies (FDPS, HMGCS1 1:1,000, Proteintech) overnight at 4°C.

Techniques: Sequencing

Journal: Frontiers in Pharmacology

Article Title: Transcriptomic Analysis Reveals the Protective Effects of Empagliflozin on Lipid Metabolism in Nonalcoholic Fatty Liver Disease

doi: 10.3389/fphar.2021.793586

Figure Lengend Snippet: Empagliflozin reduced triglyceride level both in serum (A) and liver (B) through enhanced triglyceride transfer (D) , lipolysis (E) and microsomal mitochondrial β-oxidation (F) . Empagliflozin had no effect on the synthesis genes of triglyceride (C) . Western-blot analysis showed the protein levels of FDPS and HMGCS1 (G and H) , which were the most relevant genes in lipid metabolism. Equal loading of protein was verified by probing β-actin. Data represent means ± SEM. * p < 0.05, ** p < 0.01, n = (4–6).

Article Snippet: After blocking with 5% BSA, the membranes were incubated with primary antibodies (FDPS, HMGCS1 1:1,000, Proteintech) overnight at 4°C.

Techniques: Western Blot