Journal: Molecular Biology of the Cell

Article Title: Image-based drug screen identifies HDAC inhibitors as novel Golgi disruptors synergizing with JQ1

doi: 10.1091/mbc.E17-03-0176

Figure Lengend Snippet: Image-based chemical compound screen identifies HDAC inhibitors and DNA-damaging agents as novel Golgi-dispersing compounds. To identify novel compounds that modulate Golgi morphology, a screening platform was established. (A) Screening pipeline including cell seeding (A549 cells), treatment with compound library, staining for the cis -Golgi (GM130), the nucleus (Hoechst), the ER stress marker (GRP78), cytoplasm (Phalloidin), and image acquisition and processing. (B) Representative images of the negative control (vehicle-treated cells) as well as positive controls (BFA, doxorubicin, and nocodazole) and newly discovered Golgi-fragmenting drugs (Bleomycin, Vorinostat, 4-iodo-SAHA, Trichostatin A, Givinostat, and Pracinostat) are displayed. (C) Following image analysis, to exclude potential plate effects, the Golgi area of cells treated with the chemical library was normalized to the vehicle-treated sample present within the same plate. The corresponding survival ratios of treated cells are also shown. (D) Detailed view of compounds are shown, of which at least two of three replicates caused a ≥1.5-fold increase in Golgi area. The compound panel includes positive controls and novel Golgi-fragmenting compounds, which were selected for further investigation. *** p < 0.001 vs. #; see Materials and Methods .

Article Snippet: Compounds were obtained from the following companies: brefeldin A (Sigma-Aldrich), golgicide A (Santa Cruz Biotechnology), monensin (Enzo Life Sciences), AG-1478 (Sigma), tunicamycin (Santa Cruz Biotechnology), thapsigargin (Santa Cruz Biotechnology), nocodazole (Santa Cruz Biotechnology), (+)-JQ1 (Cayman Chemical), CBP30 (TargetMol), doxorubicin (Sigma), etoposide (Sigma), teniposide (Santa Cruz Biotechnology), mitomycin-C (Santa Cruz Biotechnology), cisplatin (Santa Cruz Biotechnology), hydroxyurea (Sigma), 5-fluorouracil (Sigma), gemcitabine (Santa Cruz Biotechnology), irinotecan (Santa Cruz Biotechnology), bleomycin (Santa Cruz Biotechnology), NU7441 (Selleckchem), KU55933 (Sigma), Flavopiridol (Santa Cruz Biotechnology), phorbol 12-myristate 13-acetate (PMA; Santa Cruz Biotechnology), Panobinostat (Selleckchem), Tubastatin (Selleckchem), Entinostat (Santa Cruz Biotechnology), Pracinostat (Selleckchem), Givinostat (Selleckchem), Triptolide (Santa Cruz Biotechnology), α-Amanitin (Santa Cruz Biotechnology), and Z-VAD-FMK (Santa Cruz Biotechnology).

Techniques: Drug discovery, Staining, Marker, Negative Control

Journal: Molecules (Basel, Switzerland)

Article Title: Cytostatic Bacterial Metabolites Interfere with 5-Fluorouracil, Doxorubicin and Paclitaxel Efficiency in 4T1 Breast Cancer Cells.

doi: 10.3390/molecules29133073

Figure Lengend Snippet: Figure 9. Cadaverine and indolepropionic acid interfere with the cytostatic effect of doxorubicin. 4T1 cells were plated in 96-well plates (1500 cells/well). Cells were treated with doxorubicin alone or in combination with CAD (0.8 µM), IS (4 µM) or IPA (1 µM) for 48 h, and then cell numbers were determined by MTT assay. Data are presented as means ± SEM, from at least three biological replicates. Individual assays were measured in quadruplicate or in triplicate. Values were normalized to vehicle-treated cells (absorbance is equal to 1). Nonlinear regression (Graphpad “[Inhibitor] vs. response (four parameters)” utility) was performed on datasets to obtain IC50 and Hill slope values. Normality was determined for the inhibitory curves using the D’Agostino and Pearson normality test, while for the IC50 values and the Hill slope values the Shapiro–Wilk test was used. Statistical difference between the inhibitory curves was determined using a two-way ANOVA test, and all data points were compared with each other (in Tukey post hoc tests). For the comparison of the IC50 and Hill slope values, a non-paired, two-sided t-test was applied. ### indicates p < 0.001 for DOX-treated vs. vehicle-treated cells. * represents significance at p < 0.05 between the indicated groups. Abbreviations: CAD—cadaverine; DOX—doxorubicin; IPA—indolepropionic acid; and IS—indoxylsulfate.

Article Snippet: Liposomal Encapsuled Doxorubicin (DOX-NP, cat # 300112) was purchased from Avanti Polar Lipids (Alabaster, AL, USA) and a stock solution of 50 mM was prepared.

Techniques: MTT Assay, Comparison

Journal: bioRxiv

Article Title: The Reinstatement of a Forgotten Infantile Memory

doi: 10.1101/2025.09.27.678956

Figure Lengend Snippet: (A) Schematic of the behavioral protocol to trigger reinstatement of a forgotten infantile memory. Mice underwent contextual fear conditioning (cFC) training in infancy (P19), followed in adulthood by exposure to reminders of the CONTEXT and the AVERSIVE stimulus experienced during cFC training. The recall of the reinstated memory (REINSTATED recall) was tested in the CONTEXT one day after the final reminder. All behavioral sessions in adulthood were spaced by one day. (B) Freezing at CONTEXT reminder in adulthood was low in mice fear conditioned at P19, similar to animals that never experienced conditioning (Naϊve animals. Dunnett’s test, p = 0.634), and significantly higher in adult-conditioned animals (p < 0.001), indicating infantile amnesia. One-way ANOVA, F(2, 38) = 51.93, p < 0.001, r2 = 0.7321. n 2 11 mice per group. (C) At REINSTATED recall, mice conditioned at P19 expressed robust freezing (Dunnett’s test, p = 0.007), significantly higher than naϊve animals and comparable to animals conditioned during adulthood (> P60) only when both CONTEXT and AVERSIVE reminders were given. No reinstatement was observed in animals lacking either the CONTEXT (Dunnett’s test, p = 0.970) or the AVERSIVE (p > 0.999) reminder. One-way ANOVA, F(25, 226) = 11.18, p < 0.001, r2 = 0.5529; Brown-Forsythe test, F(25, 226) = 2.622, p < 0.001; Bartlett’s test, x2 = 81.17, p < 0.001. n 2 9 mice per group. (D) AAV-mediated expression of excitatory DREADD (hM3Dq-mCherry, yellow) in PV interneurons of dorsal hippocampus. Representative histological image showing mCherry expression in PV+ neurons in dorsal CA3 and CA1 (single-plane image, 10x objective; scale bar = 300 µm). (E, F) Activation of hippocampal PV interneurons via CNO injection during either the CONTEXT (E) or AVERSIVE (F) reminder prevented memory reinstatement, resulting in low freezing at REINSTATED recall. Student’s t -test against saline injected controls: CONTEXT, t = 3.515, p = 0.004, Cohen’s d = 0.507; AVERSIVE, t = 4.04, p = 0.003, Cohen’s d = 0.645. n = 6 for CONTEXT, n 2 4 for AVERSIVE.

Article Snippet: For bilateral delivery of DREADDs, PV-Cre mice received injections of pAAV-hSyn-DIO-hM3D(Gq)-mCherry (Addgene #44361-AAV1; titer: 2.2 x 1013 vg/mL) into the dorsal hippocampus (AP: −1.75 mm; ML: ±1.95 mm; DV: −1.85 mm; 350 nL per hemisphere) using a Nanoject III (Drummond).

Techniques: Expressing, Activation Assay, Injection, Saline

Journal: bioRxiv

Article Title: The Reinstatement of a Forgotten Infantile Memory

doi: 10.1101/2025.09.27.678956

Figure Lengend Snippet: We propose that reinstating forgotten infantile memories requires a carefully orchestrated hippocampus-centered network process that unfolds in three stages. First, exposure to a contextual reminder of the original experience primes the hippocampal network, increasing the recruitment of neurons associated with the forgotten infantile memory (iEngram) during subsequent experiences associated with the forgotten infantile memory (“ Priming ”, A ). Then, a reminder of the aversive stimulus experienced during infancy selectively tags iEngram neurons for offline reactivation in the following hours (“ Tagging ”, B ). Increased iEngram activity during offline network reactivation events binds previously latent infantile memory with novel neuronal ensembles, thereby reinstating behavioral responses consistent with the original experience (“ Binding ”, C ).

Article Snippet: For bilateral delivery of DREADDs, PV-Cre mice received injections of pAAV-hSyn-DIO-hM3D(Gq)-mCherry (Addgene #44361-AAV1; titer: 2.2 x 1013 vg/mL) into the dorsal hippocampus (AP: −1.75 mm; ML: ±1.95 mm; DV: −1.85 mm; 350 nL per hemisphere) using a Nanoject III (Drummond).

Techniques: Activity Assay, Binding Assay