rabbit polyclonal sting tmem173 antibody  (Novus Biologicals)

Journal: EMBO Reports

Article Title: DSTYK phosphorylates STING at late endosomes to promote STING signaling

doi: 10.1038/s44319-025-00394-9

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Rabbit polyclonal anti-STING , Proteintech , 19851-1-AP.

Techniques: Recombinant, Bioprocessing, Sequencing, shRNA, Transfection, Real-time Polymerase Chain Reaction, SYBR Green Assay, Mutagenesis, Software

Journal: Nature Communications

Article Title: A basally active cGAS-STING pathway limits SARS-CoV-2 replication in a subset of ACE2 positive airway cell models

doi: 10.1038/s41467-024-52803-7

Figure Lengend Snippet: a The indicated airway-derived cell lines were inoculated with WT SARS-CoV-2 (2019-nCoV/USA-WA1/2020) at a MOI: 0.15 i.u./cell. RT-qPCR analysis of cell-associated SARS-CoV-2 N sgRNA (copies/cell) at 4 and 72 hpi is shown. Data show the average of 2 independent experiments, error bars denote the range. b Focus forming assay performed on culture supernatants of the indicated cells infected with SARS-CoV-2 for 72 hpi (from panel a ). Data show focus forming units per mL (FFU/mL, log transformed) from two independent experiments, error bars show the SEM. c Polyclonal pools of ACE2 knockout (ACE2-KO) and control cell lines transduced with a non-targeting sgRNA (NT) were infected with WT SARS-CoV-2 at an MOI: 2 i.u./cell. RT-qPCR analysis of cell-associated viral RNA (sgRNA N) at 72 hpi is shown from two biological replicates. Data show the mean, error bars show the SEM. d Polyclonal pools of ACE2 KO and non-targeting control (NT) cell lines were infected with WT SARS-CoV-2 at a MOI: 2 i.u./cell. In situ hybridization of incoming viral RNA (green) at 2 hpi was performed as detailed in Methods. Nuclear DAPI staining is shown in blue. e Quantification of data presented in Panel d . Puncta of SARS-CoV-2 RNA was counted in ∼200 cells per cell line from two independent experiments. The average number of RNA puncta per condition is shown in the graph. Source data are provided as a Source Data file.

Article Snippet: After washing with PBST, samples were incubated in a goat anti-mouse secondary antibody conjugated to Alexa Fluor Plus 488 (Invitrogen, Cat# A-11029, 1:1000) at room temperature for 1 h. IFIT3, IFIT2, ISG15 and STING were detected by incubation with a primary rabbit polyclonal IFIT3 antibody (Novus Biologicals NBP2-32500, 1:500), rabbit polyclonal IFIT2 antibody (Novus Biologicals NBP2-15180SS, 1:500), rabbit polyclonal ISG15 antibody (Proteintech 15981-1-AP, 1:250) and rabbit polyclonal STING antibody (Proteintech 19851-1-AP, 1:200) respectively as described above followed by incubation with a goat anti-rabbit fluorescent secondary antibody (Invitrogen Goat anti-Rabbit Alexa Fluor Plus 568TM, Cat# A-11036, 1:1000 or Invitrogen Goat anti-Rabbit Alexa FluorTM Plus 647, Catalog # A32733TR, 1:1000).

Techniques: Derivative Assay, Quantitative RT-PCR, Focus Forming Assay, Infection, Transformation Assay, Knock-Out, Control, Transduction, In Situ Hybridization, Staining

Journal: Nature Communications

Article Title: A basally active cGAS-STING pathway limits SARS-CoV-2 replication in a subset of ACE2 positive airway cell models

doi: 10.1038/s41467-024-52803-7

Figure Lengend Snippet: a , b SCC25, H596, OE21, Detroit 562, and Calu-3 cells were pretreated with 1 μM ruxolitinib for 24 h. IFIT1 ( a ) and MX1 ( b ) expression were analyzed by RT-qPCR, and expression levels are shown relative to untreated Calu-3 cells (set to 1). Data show the mean from n = 2 (H596 and Calu-3) and 4 (other cells) independent biological replicates, error bars show the SEM. c SCC25, H596, OE21, and Detroit 562 cells were pretreated with DMSO (mock) or 1 μM ruxolitinib for 24 h and infected with SARS-CoV-2 at a MOI of 2 i.u./cell in the presence of DMSO or 1 μM ruxolitinib. RT-qPCR analysis shows cell-associated viral RNA levels at 72 hpi. Data show the mean from n = 4 independent replicates, error bars show the SEM. d–g Polyclonal populations of SCC25 cells knocked out (KO) for IFNAR ( d ) or STAT1 ( e ) using two independent sgRNAs or transduced with a non-targeting (NT) control and SCC25 cells transfected with 3 separate siRNAs targeting STAT2 ( f ), IRF9 ( g ), or a non-targeting control (NT) were infected with SARS-CoV-2 at a MOI of 2 i.u./cell. Cell-associated viral RNA was analyzed by RT-qPCR at 72 hpi. Y-axes indicate the relative expression ratio between knockdown (KD) or knockout (KO) and the respective NT samples. Data display the mean of n = 5 ( d , e ) and n = 6 ( f , g ) independent replicates, error bars denote the SEM. Source data are provided as a Source Data file.

Article Snippet: After washing with PBST, samples were incubated in a goat anti-mouse secondary antibody conjugated to Alexa Fluor Plus 488 (Invitrogen, Cat# A-11029, 1:1000) at room temperature for 1 h. IFIT3, IFIT2, ISG15 and STING were detected by incubation with a primary rabbit polyclonal IFIT3 antibody (Novus Biologicals NBP2-32500, 1:500), rabbit polyclonal IFIT2 antibody (Novus Biologicals NBP2-15180SS, 1:500), rabbit polyclonal ISG15 antibody (Proteintech 15981-1-AP, 1:250) and rabbit polyclonal STING antibody (Proteintech 19851-1-AP, 1:200) respectively as described above followed by incubation with a goat anti-rabbit fluorescent secondary antibody (Invitrogen Goat anti-Rabbit Alexa Fluor Plus 568TM, Cat# A-11036, 1:1000 or Invitrogen Goat anti-Rabbit Alexa FluorTM Plus 647, Catalog # A32733TR, 1:1000).

Techniques: Expressing, Quantitative RT-PCR, Infection, Transduction, Control, Transfection, Knockdown, Knock-Out

Journal: Nature Communications

Article Title: A basally active cGAS-STING pathway limits SARS-CoV-2 replication in a subset of ACE2 positive airway cell models

doi: 10.1038/s41467-024-52803-7

Figure Lengend Snippet: a Polyclonal populations of SCC25, H596, OE21, and Detroit 562 cells knocked out (KO) for TREX1 using two separate sgRNAs or wild-type (WT) controls were analyzed for IFIT2 expression by RT-qPCR. Data show the relative expression of IFIT2 in KO cells compared to parental controls (set to 1) from n = 3 (OE21) or 6 (SCC25, H596, Detroit 562) replicates, error bars show the SEM. b cGAS and STING variants present in the indicated cell lines derived from RNA-seq data. c , d SCC25, H596, OE21, and Detroit 562 cells grown in the presence of 10 μM VBIT-4 for 4 days ( c ) or 500 μM AZT for 3 days ( d ). IFIT1 , IFIT2, and IRF7 expression was analyzed by RT-qPCR and normalized relative to mock-treated controls (set to 1). Data show the mean from n = 3 replicates, error bars show the SEM. e Following subcellular fractionation (see “Methods”), levels of the indicated HERV and LINE DNA elements were assessed by qPCR in the cytosol and nucleus. Data show the relative levels of each element in the cytosol relative to the nuclear extract in each cell type ( n = 2, error bars show SEM). Source data are provided as a Source Data file.

Article Snippet: After washing with PBST, samples were incubated in a goat anti-mouse secondary antibody conjugated to Alexa Fluor Plus 488 (Invitrogen, Cat# A-11029, 1:1000) at room temperature for 1 h. IFIT3, IFIT2, ISG15 and STING were detected by incubation with a primary rabbit polyclonal IFIT3 antibody (Novus Biologicals NBP2-32500, 1:500), rabbit polyclonal IFIT2 antibody (Novus Biologicals NBP2-15180SS, 1:500), rabbit polyclonal ISG15 antibody (Proteintech 15981-1-AP, 1:250) and rabbit polyclonal STING antibody (Proteintech 19851-1-AP, 1:200) respectively as described above followed by incubation with a goat anti-rabbit fluorescent secondary antibody (Invitrogen Goat anti-Rabbit Alexa Fluor Plus 568TM, Cat# A-11036, 1:1000 or Invitrogen Goat anti-Rabbit Alexa FluorTM Plus 647, Catalog # A32733TR, 1:1000).

Techniques: Expressing, Quantitative RT-PCR, Derivative Assay, RNA Sequencing, Fractionation