anti eif4e  (Santa Cruz Biotechnology)

Journal: bioRxiv

Article Title: The enteroviral protease target LSM14A operates outside of P-bodies to augment antiviral innate immunity

doi: 10.64898/2026.05.11.723630

Figure Lengend Snippet: A. 293T cells were either treated with sodium arsenite (NaAsO 2 ) (100 µM for 1h at 37°C) or DMSO (vehicle), followed by staining with anti-LSM14A antibody. The graph on the right shows the number of LSM14A+ puncta in 25 randomly selected cells. Each dot represents a cell. Statistical significance was determined using two-tailed non-parametric t -test. *P < 0.05. B. 293T cells were stained with anti-LSM14A and either anti-DDX6 antibodies (Top panel) or with anti-4E-T antibodies (Bottom panel) followed by confocal microscopy. The nuclei were stained with DAPI. C. 293T cells treated with either vehicle or NaAsO 2 as in (A) were stained with anti-LSM14A and anti-G3BP1 antibodies, followed by confocal microscopy. White arrows indicate PBs juxtaposed to SGs.

Article Snippet: The antibodies used in this study include rabbit anti-LSM14A (Bethyl Laboratories; #A305-103A), mouse anti-LSM14A (Santa Cruz Biotechnology; #sc-398633), anti-DDX6 (Bethyl Laboratories; #A300-461A), anti-4E-T (Bethyl Laboratories; #A300-706A), anti-PEX19 (Proteintech; #14713-1-AP), anti-G3BP1 (Santa Cruz Biotechnology; #sc-365338), anti-PMP70 (Sigma-Aldrich; #SAB4200181), anti-HA (ABclonal; #AE008), anti-GAPDH (GeneTex; #GTX627408), and anti-β-actin (Thermo Fisher Scientific; #AM4302).

Techniques: Staining, Two Tailed Test, Confocal Microscopy

Journal: bioRxiv

Article Title: Arginine synthesis pathway and ASS1 play a critical role in mRNA translation reprogramming and ICI resistance in cutaneous melanoma

doi: 10.64898/2026.03.17.712479

Figure Lengend Snippet: A , GSEA analysis showing a set of genes significantly dysregulated in ICI-resistant cells (YUMM1.1) compared to ICI sensitive cells (YUMM2.1). B , Western blot showing indicated proteins expression levels in ICI-sensitive YUMM2.1 cells versus ICI-resistant YUMM1.1 cells. HSP90 serves as a loading control. C , Representation of the bicistronic plasmid used to compare cap-dependent and independent translation initiation (upper panel). Renilla over Firefly luminescent ratio quantification in ICI-sensitive YUMM2.1 cells versus ICI-resistant YUMM1.1 cells (n=3; mean ± SEM) (lower panel). D , Proximity Ligation Assay showing eiF4E/eiF4G interactions in YUMM2.1 and YUMM1.1 cells. Representative images are presented in the left panel, and quantifications are presented in the right panel. E-F . Representation of enriched sets of genes, obtained after GSEA analysis, in patients that progress under ICI treatment compared to patients that respond.

Article Snippet: After blocking, the antibodies were used at the following concentrations: eIF4E (mouse, clone A-10, 1:200; SantaCruz Biotechnology, sc-271480) and eIF4G (rabbit, 1:200; Cell Signaling Technology, 2498) and incubated overnight with the primary antibodies at 4 °C.

Techniques: Western Blot, Expressing, Control, Plasmid Preparation, Proximity Ligation Assay

Journal: bioRxiv

Article Title: Arginine synthesis pathway and ASS1 play a critical role in mRNA translation reprogramming and ICI resistance in cutaneous melanoma

doi: 10.64898/2026.03.17.712479

Figure Lengend Snippet: A , Western blot showing indicated proteins expression levels in YUMM1.1 cells expressing a shRNA control or targeting ASS1. HSP90 serves as a loading control. B , Western blot showing indicated proteins expression levels in YUMM1.7 cells expressing a shRNA control or targeting ASS1. HSP90 serves as a loading control. C , Western blot showing indicated proteins expression levels in YUMM2.1 cells expressing an ectopic empty vector or containing the open reading frame of ASS1. HSP90 serves as a loading control. D , Renilla over Firefly luminescent ratio quantification in YUMM1.1 cells expressing a shRNA control or targeting ASS1 (n=4; mean ± SEM). E , Renilla over Firefly luminescent ratio quantification in YUMM1.7 cells expressing a shRNA control or targeting ASS1 (n=3; mean ± SEM). F , Renilla over Firefly luminescent ratio quantification in YUMM2.1 cells expressing an ectopic empty vector or containing the open reading frame of ASS1 (n=3; mean ± SEM). G , Protein synthesis rates were determined in YUMM1.1 cells expressing a shRNA control or targeting ASS1. Cells were then pulsed for 30 min with [ 35 S]Cys/Met, and the incorporation of 35 S into proteins was quantified and normalized to the total protein amount. Cells treated 30 minutes with 100µg/mL of cycloheximide (CHX) are used as a positive control. The data are presented as the mean (n = 4). H , Protein synthesis rates were determined in YUMM1.7 cells expressing a shRNA control or targeting ASS1. Cells were then pulsed for 30 min with [ 35 S]Cys/Met, and the incorporation of 35 S into proteins was quantified and normalized to the total protein amount. Cells treated 30 minutes with 100µg/mL of cycloheximide (CHX) are used as a positive control. The data are presented as the mean (n = 3). I , Protein synthesis rates were determined in YUMM2.1 cells expressing an ectopic empty vector or containing the open reading frame of ASS1. Cells were then pulsed for 30 min with [ 35 S]Cys/Met, and the incorporation of 35 S into proteins was quantified and normalized to the total protein amount. Cells treated 30 minutes with 100µg/mL of cycloheximide (CHX) are used as a positive control. The data are presented as the mean (n = 3). J , Proximity Ligation Assay showing eiF4E/eiF4G interactions in YUMM1.1 cells expressing a shRNA control or targeting ASS1. Representative images are presented in the left panel, and quantifications are presented in the right panel. K , Proximity Ligation Assay showing eiF4E/eiF4G interactions in YUMM1.7 cells expressing a shRNA control or targeting ASS1. Representative images are presented in the left panel, and quantifications are presented in the right panel.

Article Snippet: After blocking, the antibodies were used at the following concentrations: eIF4E (mouse, clone A-10, 1:200; SantaCruz Biotechnology, sc-271480) and eIF4G (rabbit, 1:200; Cell Signaling Technology, 2498) and incubated overnight with the primary antibodies at 4 °C.

Techniques: Western Blot, Expressing, shRNA, Control, Plasmid Preparation, Positive Control, Proximity Ligation Assay