Journal: Cell reports
Article Title: Translocational attenuation mediated by the PERK-SRP14 axis is a protective mechanism of unfolded protein response.
doi: 10.1016/j.celrep.2024.114402
Figure Lengend Snippet: Figure 2. Down-regulation of SRP14 upon ER stress in multiple cells and tissues examined (A) LoVo cells expressing SP-NHK-GFP were treated with tunicamycin (TM; 5 mg/mL), thapsigargin (TG; 500 nM), or BFA (0.5 mg/mL) for 0, 4, 8, and 12 h and subjected to immunoblotting analysis with indicated antibodies. (B) Immunofluorescent staining of BiP (top) or SRP14 (bottom) together with ER-Tracker in LoVo cells treated with 500 nM TG for 8 h. The cells were then subjected to microscopic analysis. Scale bar, 25 mm. (C and D) HCT116, A549, HeLa, and SH-SY5Y cells were treated with 5 mg/mL TM for the indicated time points and subjected to immunoblotting analysis to assess the protein levels of SRP14. Quantifications of SRP14 normalized to GAPDH are show on the right.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER TnT T7 Quick for PCR DNA Promega Cat#L5540 HiPure Universal DNA Kit Magen Biotechnology Cat#IVD3018 RNAiso Plus Reagent Takara Cat#9108 Immunohistochemistry Kit (Goat Anti-Rabbit IgG H&L (HRP)) Wuhan Servicebio Technology Cat#G1215 Deposited data Whole-exome sequencing data This paper SRA: PXD052453 mRNA sequencing data This paper GEO: GSE268040 LC-MS/MS proteomics data This paper iProX: PXD052453 Experimental models: Cell lines LoVo ATCC Cat#CCL-229 LoVo-SP-NHK-GFP This paper N/A LoVo-SP-NHK-GFP Tet-ON-shHRD1 This paper N/A LoVo-2*FLAG-SRP14 This paper N/A HCT116 ATCC Cat#CCL-247 SH-SY5Y ATCC Cat#CRL-2266 A549 National Collection of Authenticated Cell Cultures Cat#TCHu150 HEK293T National Collection of Authenticated Cell Cultures Cat#GNHu44 HeLa ATCC Cat#CCL-2 HeLa-eIF2aS51A This paper N/A HeLa-PERK KO This paper N/A HeLa-ATF3 KO This paper N/A HeLa-ATF4 KO This paper N/A HeLa-FLAG-SRP14K100 This paper N/A HeLa-FLAG-SRP14K95 This paper N/A Experimental models: Organisms/strains BALB/c mice (6-week-old) Shanghai Model Organisms Center, Inc. Cat#SM-003 Oligonucleotides DNA cloning primers GENEWIZ Suzhou, China See Table S1 RT-qPCR primers GENEWIZ Suzhou, China See Table S1 siRNA & shRNA oligonucleotides Suzhou GenePharma Co., Ltd. See Table S1 sgRNA oligonucleotides GENEWIZ Suzhou, China See Table S1 EIF2S1 S51A ssODN repair template GENEWIZ Suzhou, China See Table S1 Recombinant DNA pLVX-AcGFP1-N1 Hunan Fenghui Biotechnology Co., Ltd Cat#BR020 pLVX-SP-NHK-GFP This paper N/A pLVX-PrP-GFP This paper N/A pLVX-PrPsp-NHK-GFP This paper N/A pLVX-FLAG-SP-NHK-GFP This paper N/A pLVX-FLAG-NHK-GFP This paper N/A pLVX-NHK-SP-Linker-GFP This paper N/A pLVX-Kozak-SP-NHK-GFP This paper N/A pLVX-IRES-puro Hunan Fenghui Biotechnology Co., Ltd Cat#BR025 pLVX-SP-NHK-HA This paper N/A pLVX-2*FLAG-SRP14 This paper N/A pLVX-2*FLAG-SRP54 This paper N/A (Continued on next page) 16 Cell Reports 43, 114402, July 23, 2024
Techniques: Expressing, Western Blot, Staining
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