rabbit anti ifi30  (Proteintech)

Journal: iScience

Article Title: Chronic stress antagonizes formation of stress granules

doi: 10.1016/j.isci.2025.114556

Figure Lengend Snippet: Chronic stress-pre-incubated cells fail to disassemble polysomes by acute stress U2OS cells were subjected to treatment with 100 μM SA for 1 h after pre-incubation with 10 μM SA, 1 μM Tg (thapsigargin), 60 μM CCCP (carbonyl cyanide m -chlorophenyl hydrazone), or HBSS for 24 h. Unstressed cells (NT) were used as a control. (A) Cells were pulsed with puromycin and emetine for 5 min and lysed. Cell lysates were subjected to western blotting using antibodies for puromycin, p -eIF2α, total eIF2α, G3BP1, G3BP2, caprin1, USP10, and β-actin. Three blots were taken for each of the three biological replicates (independent experiments). The same colors of the circle dot in the graph are from the same independent experiment. Results are mean ± S.E.M. ( n = 3). p values were assessed using a one-way ANOVA ( p ∗ <0.05, p ∗∗ <0.01, p ∗∗∗ <0.001, p ∗∗∗∗ <0.0001). (B–F) Polysome profiles from U2OS cells. NT, black; 1 h SA, red; (B) 24 h pre-incubation of SA, blue; 1 h SA with 24 h pre-incubation of SA, green. (C) 24 h pre-incubation of Tg, blue; 1 h SA with 24 h pre-incubation of Tg, green. (D) 24 h pre-incubation of CCCP, blue; 1 h SA with 24 h pre-incubation of CCCP; green. (E) 24 h pre-incubation of HBSS, blue; 1 h SA with 24 h pre-incubation of HBSS; green. (F) The polysome/monosome ratio was calculated with normalization by each of NT as a control.

Article Snippet: Rabbit Polyclonal Caprin1 antibody , Proteintech , 15112-1-AP; RRID: AB_2070016.

Techniques: Incubation, Control, Western Blot

Journal: Cell Reports Methods

Article Title: Combination of in vivo and in vitro phosphoproteomics determines the PP2A target repertoire on proteome scale

doi: 10.1016/j.crmeth.2025.101084

Figure Lengend Snippet: Characterization of bona fide PP2A-PPP2R5E target sites (A) Complexome profiling of StrepHA-PPP2R5E purifications indicates enrichment of holocomplexes. (B) Volcano plot highlighting identified phosphatase subunits in StrepHA- PPP2R5E (left) and StrepHA-PPP1CA (right) purifications (marked in red) under native conditions. Dotted lines mark significance threshold, significantly enriched proteins marked in blue ( n = 3 biological replicates, FDR <0.05). Significantly enriched phosphatase subunits are annotated. (C) Significantly regulated phosphosites for PPP2R5E-PP2A complexes being OA sensitive ( n = 3 biological replicates). Significant sites are highlighted in blue (FDR <0.05). (D) Comparison of OBIPhA and in vivo analyses. Venn diagram highlights the overlap of identified phosphosites using the different experimental approaches. Bona fide PPP2R5E-PP2A target sites are defined as being regulated in vivo and in vitro (i.e., 194 sites on 168 proteins). (E) SLiMs in the bona fide PPP2R5E/B56ε-PP2A targets. (F) GO enrichment analysis using STRING DB highlighting biological processes of PPP2R5E-PP2A target proteins. (G) Targeted, phosphosite-specific MS analysis (PRM) for DDX3X Ser90 and Ser609. Shown is the quantification of three replicates (black dots). ∗ p < 0.05; ∗∗ p < 0.01; t test. (H) IF of CAPRIN1 and G3BP1. Shown are exemplary images of n = 3 biological replicates. Nuclei are stained in blue. Scale bar, 10 μm. Box plots show quantifications of 12 images of n = 3 biological replicates, boxes are drawn according to ggplot2 standard settings. White dots indicate average stress granule number/mean volume per cell and image. ∗ p < 0.05; ∗∗ p < 0.01; t test. See also .

Article Snippet: Rabbit polyclonal anti Caprin1 , ProteinTech , Cat#15112-1-AP RRID: AB_2070016.

Techniques: Comparison, In Vivo, In Vitro, Phospho-proteomics, Staining