Journal: Vaccines

Article Title: In-Use Stability and Device Compatibility Define Clinically Actionable Handling Limits for a GMP-Produced Attenuated Listeria monocytogenes Vaccine Expressing GUCY2C

doi: 10.3390/vaccines14050461

Figure Lengend Snippet: Lm-GUCY2C Functional Testing. J774A.1 cells were infected in duplicate with vehicle, parental Lm, or Lm-GUCY2C. Cell lysates were then examined by immunoblot for GUCY2C and LLO.

Article Snippet: Murine macrophage-like J774A.1 cells (ATCC TIB-67) were maintained in DMEM supplemented with 10% FBS and penicillin/streptomycin (100 U/mL; 100 μg/mL) at 37 °C, 5% CO 2 .

Techniques: Functional Assay, Infection, Western Blot

Journal: Journal of Traditional and Complementary Medicine

Article Title: Cinnamaldehyde inhibits the NLRP3 inflammasome and inflammatory response by reducing oxidative stress in Neisseria gonorrhoeae-infected macrophages

doi: 10.1016/j.jtcme.2025.06.003

Figure Lengend Snippet: Effect of CA on the NLRP3 inflammasome in N. gonorrhoeae -infected macrophages. (A – D) J774A.1 macrophages were incubated with CA (5–20 μM) or vehicle for 0.5 h before N. gonorrhoeae infection for an additional 24 h. The levels of pro-caspase-1 (p45)/active caspase-1 (p10) (A ), IL-1β (C ) and IL-18 (D ) in the supernatants were analysed by Western blotting. The levels of IL-1β in the supernatants were analysed by ELISA (B) . (E) PBMCs were incubated with CA (5–20 μM) or vehicle for 0.5 h before N. gonorrhoeae infection for an additional 24 h. The levels of IL-1β in the supernatants were analysed by ELISA. (F) J774A.1 macrophages were incubated with CA (20–160 μM) or vehicle for 24 h. The levels of LDH in the supernatants were analysed by an LDH release assay kit. The ELISA and LDH release data are expressed as the mean ± SD of three separate experiments. The Western blotting images are representative of different experiments, and the accompanying histogram illustrates the quantification achieved by analyzing band intensity using ImageJ software, expressed as the mean ± SD for these three experiments. ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001 compared to N. gonorrhoeae -infected macrophages or control cells.

Article Snippet: The mouse macrophage cell line J774A.1 was obtained from the American Type Culture Collection (Rockville, MD).

Techniques: Infection, Incubation, Western Blot, Enzyme-linked Immunosorbent Assay, Lactate Dehydrogenase Assay, Software, Control

Journal: Journal of Traditional and Complementary Medicine

Article Title: Cinnamaldehyde inhibits the NLRP3 inflammasome and inflammatory response by reducing oxidative stress in Neisseria gonorrhoeae-infected macrophages

doi: 10.1016/j.jtcme.2025.06.003

Figure Lengend Snippet: Effect of CA on pyroptosis in N. gonorrhoeae -infected macrophages. J774A.1 macrophages were incubated with CA (5–20 μM) or vehicle for 0.5 h before N. gonorrhoeae infection for an additional 24 h. The levels of caspase-11 (A) , GSDMD (C) , NLRP3 (E) , ASC (F) in the supernatants, and GSDMD in the cell lysates (B) were analysed by Western blotting. (D) The levels of LDH in the supernatants were analysed by an LDH release assay kit. The LDH release data are expressed as the mean ± SD of three separate experiments. The Western blotting images are representative of different experiments, and the accompanying histogram illustrates the quantification achieved by analyzing band intensity using ImageJ software, expressed as the mean ± SD for these three experiments. ∗ p < 0.05 compared to N. gonorrhoeae -infected macrophages.

Article Snippet: The mouse macrophage cell line J774A.1 was obtained from the American Type Culture Collection (Rockville, MD).

Techniques: Infection, Incubation, Western Blot, Lactate Dehydrogenase Assay, Software

Journal: Journal of Traditional and Complementary Medicine

Article Title: Cinnamaldehyde inhibits the NLRP3 inflammasome and inflammatory response by reducing oxidative stress in Neisseria gonorrhoeae-infected macrophages

doi: 10.1016/j.jtcme.2025.06.003

Figure Lengend Snippet: Effect of CA on proinflammatory mediator expression in N. gonorrhoeae -infected macrophages. J774A.1 macrophages were incubated with CA (5–20 μM) or vehicle for 0.5 h before N. gonorrhoeae infection for an additional 24 h. The levels of IL-6 (A) and TNF-α (C) in the supernatants were analysed by ELISA. The levels of iNOS (B) , COX-2 (D) , pro-IL-1β (E) and NLRP3 (F) in the cell lysates were analysed by Western blotting. The ELISA data are expressed as the mean ± SD of three separate experiments. The Western blotting images are representative of different experiments, and the accompanying histogram illustrates the quantification achieved by analyzing band intensity using ImageJ software, expressed as the mean ± SD for these three experiments. ∗ p < 0.05, ∗∗ p < 0.05 and ∗∗∗ p < 0.001 compared to N. gonorrhoeae -infected macrophages.

Article Snippet: The mouse macrophage cell line J774A.1 was obtained from the American Type Culture Collection (Rockville, MD).

Techniques: Expressing, Infection, Incubation, Enzyme-linked Immunosorbent Assay, Western Blot, Software

Journal: Journal of Traditional and Complementary Medicine

Article Title: Cinnamaldehyde inhibits the NLRP3 inflammasome and inflammatory response by reducing oxidative stress in Neisseria gonorrhoeae-infected macrophages

doi: 10.1016/j.jtcme.2025.06.003

Figure Lengend Snippet: Effect of CA on MAPK phosphorylation and NF-κB activation in N. gonorrhoeae -infected macrophages. J774A.1 macrophages were incubated with CA (5–20 μM) or vehicle for 0.5 h before N. gonorrhoeae infection for an additional 4 h. The phosphorylation levels of ERK1/2 (A) , JNK1/2 (B), p38 (C) , IKKα/β (D) and IκBα (E) in the cell lysates were analysed by Western blotting. (F) J-Blue cells were incubated with CA (5–20 μM) or vehicle for 0.5 h before N. gonorrhoeae infection for an additional 24 h. The transcriptional activity of NF-κB was analysed by an NF-κB reporter assay. The Western blotting images are representative of different experiments, and the accompanying histogram illustrates the quantification achieved by analyzing band intensity using ImageJ software, expressed as the mean ± SD for these three experiments. The NF-κB reporter assay results are expressed as the mean ± SD of three separate experiments. ∗ p < 0.05 compared to N. gonorrhoeae -infected macrophages.

Article Snippet: The mouse macrophage cell line J774A.1 was obtained from the American Type Culture Collection (Rockville, MD).

Techniques: Phospho-proteomics, Activation Assay, Infection, Incubation, Western Blot, Activity Assay, Reporter Assay, Software

Journal: Journal of Traditional and Complementary Medicine

Article Title: Cinnamaldehyde inhibits the NLRP3 inflammasome and inflammatory response by reducing oxidative stress in Neisseria gonorrhoeae-infected macrophages

doi: 10.1016/j.jtcme.2025.06.003

Figure Lengend Snippet: Effect of CA on intracellular H 2 O 2 and mitochondrial damage in N. gonorrhoeae -infected macrophages. (A) J774A.1 macrophages were incubated with 20 μM CA or vehicle for 0.5 h before N. gonorrhoeae infection for an additional 4 h. The levels of intracellular H 2 O 2 were analysed by DCFH 2 -DA staining. (B – D) J774A.1 macrophages were incubated with 20 μM CA or vehicle for 0.5 h before N. gonorrhoeae infection for an additional 24 h. (B) The levels of mitochondrial ROS were analysed by MitoSOX staining. (C) Mitochondrial membrane integrity was analysed by MitoTracker Deep Red and MitoTracker Green staining. (D) The mitochondrial membrane potential was analysed by DiOC 2 (3) staining. The data are expressed as the mean ± SD of three separate experiments. ∗ p < 0.05 as indicated.

Article Snippet: The mouse macrophage cell line J774A.1 was obtained from the American Type Culture Collection (Rockville, MD).

Techniques: Infection, Incubation, Staining, Membrane