primary antibody cgas  (Proteintech)

Journal: The Journal of Biological Chemistry

Article Title: MYC controls STING levels to downregulate inflammatory signaling in breast cancer cells upon DNA damage

doi: 10.1016/j.jbc.2025.108560

Figure Lengend Snippet: MYC suppresses irradiation or etoposide-induced chemokine production. A , representative immunofluorescence images of BT-549 cells, either left untreated (control) or irradiated (5 Gy), stained with DAPI ( blue ) and anti-cGAS ( green ). Scale bar represents 10 μM. Quantification of cGAS-positive micronuclei is shown, with error bars representing the SD of three independent experiments. B , representative immunofluorescence images of BT-549 cells, either left untreated (control) or treated with etoposide (1 μM) BT-549 for 48 h; cells at different time points. Scale bar represents 5 μM. Quantification of cGAS-positive micronuclei is indicated for multiple treatment periods, with error bars representing the SD of three independent experiments. C , BT-549 cells were irradiated (3 Gy) and immunoblotted for pSTAT1, pTBK1, RELB, and STING, and qPCR analysis was done for CCL5 and ISG15 expression. Error bars represent the SD of four independent replicates. D , BT-549 STING WT and STING KO cells were treated with etoposide (3 Gy) and immunoblotted for pSTAT1, pTBK1, RELB, and STING, and qPCR analysis was done for CCL5 and ISG15 expression. Data of at least three independent replicates are plotted. E and F , qPCR analysis of MYC and CCL5 expression in MYC WT ( blue ) and MYC L420P ( yellow ) overexpressing cells after irradiation (IR) (n = 5, panel E) or etoposide (eto) treatment (n = 4, panel F). Throughout the figure, statistical analysis was done using unpaired two-tailed Student’s t-tests.

Article Snippet: Subsequently, cells were permeabilized with 0.1% Triton X-100 in PBS for 1 minute followed by blocking in 0.05% Tween-20 and 2.5% BSA in PBS for 1 h. Cells were incubated overnight with primary antibodies against cGAS (1:200, Cell Signaling, #15102) in PBS–Tween–BSA.

Techniques: Irradiation, Immunofluorescence, Control, Staining, Expressing, Two Tailed Test

Journal: Drug Development Research

Article Title: Dihydroartemisinin Attenuates Radiation‐Induced Lung Injury by Inhibiting the cGAS/STING/NF‐κB Signaling Pathway

doi: 10.1002/ddr.70090

Figure Lengend Snippet: DHA inhibited the expression of cGAS/STING/NF‐κB signaling pathway in lung tissue. (A) Transcriptome analysis shown that there were significant changes in gene expression between the control group and the IR group. (B) Dotplot shown KEGG representative top20 pathway entries enriched in mouse lung tissue. (C) The affinity between DHA and cGAS was assayed through molecular docking; The affinity of the H‐bond (TYR‐346, HIS‐369) was −7.2 kcal/mol. (D) Representative cGAS and STING IHC staining and semi‐quantitative analysis were performed in mouse lung tissues. The cGAS‐positive area ratios and the STING‐positive area ratios. scale: 200 μm (×100) and 20 μm (×400). (E) Western blot of cGAS, STING, NF‐κB and p‐NF‐κB proteins. Corresponding quantification of various proteins, and β‐actin as the loading control. DHA, dihydroartemisinin; IR, irradiation; RT, radiotherapy. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: For IHC, the paraffin sections were incubated overnight at 4°C with primary antibodies against cGAS (1:1000, Cell Signaling Technology), STING (1:1000, Cell Signaling Technology), TGF‐β (1:100, Servicebio), IL‐6 (1:500, Servicebio) and TNF‐α (1:500, Servicebio), succeeded by incubation at 37°C for 60 min with mouse secondary antibody (1:200; Servicebio).

Techniques: Expressing, Gene Expression, Control, Immunohistochemistry, Western Blot, Irradiation

Journal: Drug Development Research

Article Title: Dihydroartemisinin Attenuates Radiation‐Induced Lung Injury by Inhibiting the cGAS/STING/NF‐κB Signaling Pathway

doi: 10.1002/ddr.70090

Figure Lengend Snippet: DHA inhibited the expression of cGAS/STING/NF‐κB signaling pathway in BEAS‐2B cells. (A) Western blot of cGAS, STING, NF‐κB and p‐NF‐κB proteins. Corresponding quantification of various proteins, and β‐actin as the loading control. (B) Relative mRNA expression of cGAS and STING in BEAS‐2B cells after irradiation. DHA, dihydroartemisinin; IR, irradiation. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: For IHC, the paraffin sections were incubated overnight at 4°C with primary antibodies against cGAS (1:1000, Cell Signaling Technology), STING (1:1000, Cell Signaling Technology), TGF‐β (1:100, Servicebio), IL‐6 (1:500, Servicebio) and TNF‐α (1:500, Servicebio), succeeded by incubation at 37°C for 60 min with mouse secondary antibody (1:200; Servicebio).

Techniques: Expressing, Western Blot, Control, Irradiation

Journal: Drug Development Research

Article Title: Dihydroartemisinin Attenuates Radiation‐Induced Lung Injury by Inhibiting the cGAS/STING/NF‐κB Signaling Pathway

doi: 10.1002/ddr.70090

Figure Lengend Snippet: Downregulating cGAS further enhances the alleviating effects of DHA on radiation‐induced lung injury. (A) Relative mRNA expression and (B) Western blot of cGAS in BEAS‐2B cells after siRNA transfection. (C) The cell viability of BEAS‐2B cells transfected with siRNA after 8 Gy irradiation was detected by CCK‐8. (D) Western blot of cGAS, STING, NF‐κB and p‐NF‐κB proteins after siRNA transfection. Corresponding quantification of various proteins, and β‐actin as the loading control. (E) Relative mRNA expression of cGAS, STING, TGF‐β, IL‐6 and TNF‐α in BEAS‐2B cells after siRNA transfection. (F) The levels of TGF‐β, IL‐6 and TNF‐α in the culture medium of BEAS‐2B cells after siRNA transfection were detected by ELISA. CCK‐8, Cell Counting Kit‐8; DHA, dihydroartemisinin; IR, irradiation; siRNA, small interfering RNA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: For IHC, the paraffin sections were incubated overnight at 4°C with primary antibodies against cGAS (1:1000, Cell Signaling Technology), STING (1:1000, Cell Signaling Technology), TGF‐β (1:100, Servicebio), IL‐6 (1:500, Servicebio) and TNF‐α (1:500, Servicebio), succeeded by incubation at 37°C for 60 min with mouse secondary antibody (1:200; Servicebio).

Techniques: Expressing, Western Blot, Transfection, Irradiation, CCK-8 Assay, Control, Enzyme-linked Immunosorbent Assay, Cell Counting, Small Interfering RNA