Journal: Cell

Article Title: RNA Pol II inhibition activates cell death independently from the loss of transcription

doi: 10.1016/j.cell.2025.07.034

Figure Lengend Snippet: (A) TRP-induced death in U2OS and PTBP1-KO. Data are mean ± SD for 6 biological replicates. (B) PTBP1-dependence for RNA Pol II degraders, or canonical apoptotic agents: 1 μM TRP, 10 μM ⍺-amanitin (⍺-ama.), 100 μM ABT-199 (ABT), and 0.5 μM staurosporine (STS). Drugs tested in U2OS, PTBP1-KO, or BAX/BAK DKO. LF kinetics with area under the curve colored: gray, WT; red, DKO or PTBP1-KO, as indicated. Areas based on mean of 6 biological replicates. (C) RNA-seq comparing drug-induced transcriptional changes in U2OS (WT) and PTBP1-KO, 8 h after TRP. Data normalized using ERCC spike-ins. Dashed line, x = y. Pearson correlation coefficient shown. Anti-apoptotic (anti-APO) and pro-apoptotic (pro-APO) regulators highlighted. (D) Chemogenetic profiling data, highlighting genes alternatively spliced in a TRP/PTBP1-KO dependent manner. Blue, increased in PTBP-KO; red, decreased in PTBP1-KO. See also . (E) CoIP of RNA Pol IIA with mCherry-PTBP1. (F) CoIP of PTBP1 with varied forms of RNA Pol II. (G) Localization of mCherry-PTBP1 ± TRP for 48 h. Scale bar, 25 μm. (H) Quantification of PTBP1 cytoplasm/nuclear ratio following TRP. (I and J) Fractionation to assess PTBP1 localization following exposure to ⍺-ama. W, whole-cell lysate; C, cytoplasmic; N, nuclear. (I) U2OS-RBP1-N792D/ΔCTD cells following 12-h 10 μM ⍺-amanitin. (J) As in (I), except with doxycycline (+Dox). See also and and .

Article Snippet: Anti-mCherry rabbit polyclonal Ab , Proteintech , Cat# 26765–1-AP; RRID: AB_2876881.

Techniques: RNA Sequencing, Fractionation

Journal: Cell

Article Title: RNA Pol II inhibition activates cell death independently from the loss of transcription

doi: 10.1016/j.cell.2025.07.034

Figure Lengend Snippet: (A) Strategy to identify PDAR-dependent drugs. KO refers to genetic dependencies observed for RNA Pol II degraders. (B) Cell death in U2OS for a high-scoring drug (10 μM ⍺-ama) and low-scoring drug (3.16 μM STS). Data are mean ± SD of 3 biological replicates. (C) Heatmap depicting PDS score across a 7-point, half-log dose range for 46 compounds. Data collected in U2OS. Gray boxes, non-lethal doses. ***Clinically approved; *clinically investigated. (D) Binary classifier, ordered as in (C). Red circles denote PD-like mechanisms. (E) mCherry-PTBP1 cytoplasmic-nuclear (Cyto/Nuc.) ratio following indicated compounds. (F) Immunoblot of Rpb1 in U2OS following 1 μM abemaciclib (PD-like) or a 10-fold higher dose (low PDS). (G) As in (F), but for cisplatin. (H) PDS across a panel of cell lines for 1 μM TRP, 10 μM flavopiridol, 100 μM cisplatin, and 100 μM ABT-199. (I) Relationship between LF 50 and RNA Pol IIA t 1/2 . Data in gray denote established transcriptional inhibitors, as shown in . Red and blue data represent PD-like and non-PD-like compounds, respectively. See also and .

Article Snippet: Anti-mCherry rabbit polyclonal Ab , Proteintech , Cat# 26765–1-AP; RRID: AB_2876881.

Techniques: Western Blot

Journal: Frontiers in Neuroscience

Article Title: Chemogenetic tools for modulation of spatial learning in dopamine transporter deficient rats

doi: 10.3389/fnins.2025.1615208

Figure Lengend Snippet: Immunofluorescence staining of rat brain LC neurons after administration of CAV PRS hM3D(Gq)-mCherry virus to the PFC (A,B) ; green fluorescence is DβH-positive neurons (C) , red fluorescence is mCherry-positive neurons (D) , double-labeled neurons appear yellow-orange (E) .

Article Snippet: Then, the sections were incubated in the primary antibodies: mouse anti-DβH antibody (1:2000, MAB308, Chemicon) and rabbit anti-mCherry antibody (1:1000 Cat#632496, Takara Bio, United States) ( ) for 24 h at RT, and the secondary antibody: donkey anti-mouse IgG Alexa Fluor 488 (1:500, abcam, ab150105, UK) and CyTM3 AffiniPure donkey anti-rabbit IgG (1:800, AB_2307443, Jackson ImmunoResearch Labs, United States) for 2 h at 37°C.

Techniques: Immunofluorescence, Staining, Virus, Fluorescence, Labeling