sting inhibitor h151  (MedChemExpress)

Journal: Cell Death & Disease

Article Title: Molecular features of TNBC govern heterogeneity in the response to radiation and autophagy inhibition

doi: 10.1038/s41419-025-07873-w

Figure Lengend Snippet: Cells were treated with the STING inhibitor H151 (10 μM), the TBK1/IKKε inhibitor amlexanox (20 μM) or the NF-κB inhibitor BI605906 (10 μM) alone or in combination with A 3×8 Gy X-rays in MDA-MB-468, B 8 Gy C-ions and 3 μM SAR405 in MDA-MB-436 and C 3 μM SAR405 in MDA-MB-231. H151, amlexanox and BI605906 were applied directly after the last irradiation/SAR405 treatment and kept for 72 hours. All cells were harvested after 72 hours of treatment. RT-qPCR analysis of IFNB1 , CXCL10 , PD-L1 and IL-6 mRNA levels is shown. TBP served as a housekeeping gene. The bars and error bars represent mean values ± standard deviation. Three biologically independent experiments were performed. P-values were calculated with a one-way ANOVA (* ≤ 0.05; **≤ 0.01; ***≤ 0.005, ****≤0.0001).

Article Snippet: The STING inhibitor H151 (TargetMol, T5674) and the NF-κB inhibitor BI-605906 (THP Medical Products, HY-13019) were used at 10 μM.

Techniques: Irradiation, Quantitative RT-PCR, Standard Deviation

Journal: bioRxiv

Article Title: α-Synuclein aggregates induce mitochondrial damage and trigger innate immunity to drive neuron–microglia communication

doi: 10.1101/2025.06.23.661105

Figure Lengend Snippet: 𝜶 -Syn spread to microglia via TNTs. (a) Representative confocal images of neuronal cells loaded with α-Syn (donors) in co-culture for 24h with microglial cells (acceptors) in the presence of the STING activator diABZI alone, or alongside the STING inhibitor H-151. Cyan asterisks highlight acceptor microglial cells positive for α- Syn. (b) Quantification of the percentage of acceptor microglial cells positive for α-Syn. N=3 independent experiments, n=25-26 fields of views. Statistical significance was analyzed using Kruskal Wallis test with Dunn’s multiple comparison. ns: p>0.05, ***p<0.001. (c) Quantification of the number of α-Syn aggregates per acceptor microglial cell as in (b). n=106 cells for control, 177 cells for diABZI-treated group, and 92 cells for diABZI+H-151 co-treated group. Statistical significance was analyzed using Kruskal Wallis test with Dunn’s multiple comparison. ns: p>0.05, *p<0.05. (d) Quantification of the extent of α-Syn transfer via secretion (CM: conditioned media). N=3 independent experiments, n=15 fields of views. Statistical significance was analyzed using Kruskal Wallis test with Dunn’s multiple comparison. ns: p>0.05. (e) Representative confocal images of neuronal cells loaded with α- Syn (donors) in co-culture for 24h with microglial cells (acceptors) in the presence of pro- inflammatory cytokines alone (upper panels) or co-treated with the NF-κB inhibitor JSH-23 (lower panels). Cyan asterisks highlight acceptor microglial cells positive for α-Syn. (f) Quantification of the percentage of acceptor microglial cells positive for α-Syn. N=3 independent experiments, n=24-25 fields of views. Statistical significance was analyzed using Two-Way ANOVA with Šídák’s multiple comparison. *p<0.05, **p<0.01, ****p<0.0001. (g) Quantification of the number of α-Syn aggregates per acceptor microglial cell as in (f). n=126 (Control –JSH-23), 124 (control +JSH-23), 227 (IL-1α –JSH-23), 163 (IL-1α +JSH-23), 223 (IL-1𝛽 –JSH-23), 144 (IL-1𝛽 +JSH-23), 188 (TNFα –JSH-23), and 134 (TNFα +JSH-23) cells. Statistical significance was analyzed using Two-Way ANOVA with Šídák’s multiple comparison. ns: p>0.05, ***p<0.001, ****p<0.0001. (h) Quantification of the extent of α-Syn transfer via secretion (CM: conditioned media). N=3 independent experiments, n=14-16 fields of views. Statistical significance was analyzed using Two-Way ANOVA with Šídák’s multiple comparison. ns: p>0.05, *p<0.05. Data in all the graphs are represented as median and quartiles. Mean values are mentioned within the graphs.

Article Snippet: For inflammatory modulations, following reagents were used with associated treatment concentration and durations: STING activator – diABZI (SelleckChem – compound 3, S8796; 2.5 μM for 2h towards the end of experiment), STING inhibitor – H-151 (Invivogen InvitroFit TM , inh-h151; 0.5 μg/mL for 16h for TNT assessment experiments, and 24h for α-Syn transfer experiments), NF-κB inhibitor – JSH-23 (Sigma, J4455; 10 μM for 16h for TNT assessment experiments, and 24h for α-Syn transfer experiments), human cGAS inhibitor – G140 (Invivogen InvitroFit TM , inh-g140; 10 μM for 16h), Bcl-2 inhibitor – ABT-737 (Sigma, 197333; 10 μM for 6h, 12h, or 24h) co-treated with pan-caspase inhibitor – Q-VD-OPh (Non-O- methylated, Sigma, 551476, 10 μM), human IL-1α (Sigma, I2778; 3 ng/mL for 24h), human IL-1β (Sigma, H6291; 20 ng/mL for 24h), human TNF-α (Merck Millipore, 654205; 50 ng/mL for 24h), TLR4 inhibitor – TAK-242 (Tocris, 6587; 10 μM for 16h), TLR2 inhibitor – TL2-C29 (Invivogen InvitroFit TM , inh-C29; 100 μM for 16h), lysosomotropic agent LLOMe (Sigma, L7393; 1 mM for 1h towards the end of experiment), mitochondrial electron transport chain uncoupler CCCP (Sigma, C2759; 20 μM for 2h).

Techniques: Co-Culture Assay, Comparison, Control

Journal: bioRxiv

Article Title: α-Synuclein aggregates induce mitochondrial damage and trigger innate immunity to drive neuron–microglia communication

doi: 10.1101/2025.06.23.661105

Figure Lengend Snippet: (a) Representative confocal images of mitochondria-labeled control (upper panels) or α-Syn-loaded (bottom panels) neuronal cells (donors) in co-culture for 24h with microglial cells (acceptors). (b) Quantification of the percentage of microglial cells receiving MitoDsRed+ mitochondria in both co-cultures and conditioned media (secretion) experiments. N=3 independent experiments, n=35 (N MitoDsRed + M co-culture), 47 (N MitoDsRed α-Syn + M co- culture), 21 (N MitoDsRed + M secretion) and 22 (N MitoDsRed α-Syn + M secretion) fields of views. Statistical significance was analyzed using Two-Way ANOVA with Uncorrected Fisher’s LSD multiple comparison. ns: p>0.05, ****p<0.0001. (c) Quantification of the number of mitochondrial particles received per acceptor microglia as in (b). N=74 (N MitoDsRed + M co- culture), 185 (N MitoDsRed α-Syn + M co-culture), 30 (N MitoDsRed + M secretion), and 34 (N MitoDsRed α- Syn + M secretion) cells. Statistical significance was analyzed using Two-Way ANOVA with Uncorrected Fisher’s LSD multiple comparison. ns: p>0.05, ***p<0.001. (d) Representative confocal images of neuronal cell-derived MitoDsRed+ mitochondria in microglial cells. (e-f) Quantification of the proportion of transferred mitochondria from naïve neuronal cells (e) or α- Syn-burdened neuronal cells (f) that remain isolated in microglial cells, or overlap with the host network. Total number of mitochondrial particles analyzed is mentioned within the pie-chart. (g) Quantification of the relative health of transferred mitochondria (Cytochrome c level, indicative of outer membrane permeabilization state of mitochondria) from naïve (N MitoDsRed + M) or α-Syn-burdened (N MitoDsRed α-Syn + M) neuronal cells. N=3 independent experiments, n=50 transferred mitochondrial particles analyzed. Statistical significance was tested using Mann-Whitney test. *p<0.05. (h) Representative confocal image, and orthogonal views of dashed boxes, highlighting encapsulation of MitoDsRed+ mitochondrial particles derived from α-Syn-burdened neuronal cells within LAMP1+ lysosomes of microglial cells. (i) Quantification of overlap between MitoDsRed+ mitochondria and LAMP1+ lysosomes in acceptor microglial cells. Data in violin plots are represented as median and quartiles, with mean values mentioned within the graphs.

Article Snippet: For inflammatory modulations, following reagents were used with associated treatment concentration and durations: STING activator – diABZI (SelleckChem – compound 3, S8796; 2.5 μM for 2h towards the end of experiment), STING inhibitor – H-151 (Invivogen InvitroFit TM , inh-h151; 0.5 μg/mL for 16h for TNT assessment experiments, and 24h for α-Syn transfer experiments), NF-κB inhibitor – JSH-23 (Sigma, J4455; 10 μM for 16h for TNT assessment experiments, and 24h for α-Syn transfer experiments), human cGAS inhibitor – G140 (Invivogen InvitroFit TM , inh-g140; 10 μM for 16h), Bcl-2 inhibitor – ABT-737 (Sigma, 197333; 10 μM for 6h, 12h, or 24h) co-treated with pan-caspase inhibitor – Q-VD-OPh (Non-O- methylated, Sigma, 551476, 10 μM), human IL-1α (Sigma, I2778; 3 ng/mL for 24h), human IL-1β (Sigma, H6291; 20 ng/mL for 24h), human TNF-α (Merck Millipore, 654205; 50 ng/mL for 24h), TLR4 inhibitor – TAK-242 (Tocris, 6587; 10 μM for 16h), TLR2 inhibitor – TL2-C29 (Invivogen InvitroFit TM , inh-C29; 100 μM for 16h), lysosomotropic agent LLOMe (Sigma, L7393; 1 mM for 1h towards the end of experiment), mitochondrial electron transport chain uncoupler CCCP (Sigma, C2759; 20 μM for 2h).

Techniques: Labeling, Control, Co-Culture Assay, Comparison, Derivative Assay, Isolation, Membrane, MANN-WHITNEY, Encapsulation

Journal: bioRxiv

Article Title: α-Synuclein aggregates induce mitochondrial damage and trigger innate immunity to drive neuron–microglia communication

doi: 10.1101/2025.06.23.661105

Figure Lengend Snippet: (a) Representative super-resolution SIM images of α-Syn-burdened MitoDsRed+ neuronal cells in co-culture with microglial cells for 24h. Insets i, ii, and iii highlight transferred mitochondrial particles that are mtDNA+, mtDNA-, and have mtDNA extruding (white arrowhead) from the matrix, respectively. (b) Quantification of the proportion of transferred mitochondria from control or α-Syn-burdened neuronal cells that are positive or negative for mtDNA. N=3 independent experiments, numbers of mitochondrial particles analyzed are mentioned within the bars. Error bars represent SEM. (c) Representative confocal images of STING translocation to GM130+ Golgi in microglial cells when co-cultured for 24h with naïve (left panels) or α-Syn-burdened (right panels) neuronal cells. (d) Quantification of mean STING fluorescence intensity on GM130+ Golgi in microglial cells. N=3 independent experiments, n=100 cells. Statistical significance was analyzed using Mann-Whitney test. **p<0.01. (e) Representative confocal images of nuclear translocation of NF-κB in microglial cells when co- cultured for 24h with naïve (left panels) or α-Syn-burdened (right panels) neuronal cells. (f) Quantification of mean nuclear NF-κB fluorescence intensity in microglial cells. N=3 independent experiments, n=100 cells. Statistical significance was analyzed using Mann- Whitney test. ****p<0.0001. (g) Representative confocal images of nuclear translocation of IRF3 in microglial cells when co-cultured for 24h with naïve (left panels) or α-Syn-burdened (right panels) neuronal cells. (h) Quantification of mean nuclear IRF3 fluorescence intensity in microglial cells. N=3 independent experiments, n=100 cells. Statistical significance was analyzed using Mann-Whitney test. ****p<0.0001. Data in violin graphs are represented as median and quartiles. Mean values are mentioned within the graphs.

Article Snippet: For inflammatory modulations, following reagents were used with associated treatment concentration and durations: STING activator – diABZI (SelleckChem – compound 3, S8796; 2.5 μM for 2h towards the end of experiment), STING inhibitor – H-151 (Invivogen InvitroFit TM , inh-h151; 0.5 μg/mL for 16h for TNT assessment experiments, and 24h for α-Syn transfer experiments), NF-κB inhibitor – JSH-23 (Sigma, J4455; 10 μM for 16h for TNT assessment experiments, and 24h for α-Syn transfer experiments), human cGAS inhibitor – G140 (Invivogen InvitroFit TM , inh-g140; 10 μM for 16h), Bcl-2 inhibitor – ABT-737 (Sigma, 197333; 10 μM for 6h, 12h, or 24h) co-treated with pan-caspase inhibitor – Q-VD-OPh (Non-O- methylated, Sigma, 551476, 10 μM), human IL-1α (Sigma, I2778; 3 ng/mL for 24h), human IL-1β (Sigma, H6291; 20 ng/mL for 24h), human TNF-α (Merck Millipore, 654205; 50 ng/mL for 24h), TLR4 inhibitor – TAK-242 (Tocris, 6587; 10 μM for 16h), TLR2 inhibitor – TL2-C29 (Invivogen InvitroFit TM , inh-C29; 100 μM for 16h), lysosomotropic agent LLOMe (Sigma, L7393; 1 mM for 1h towards the end of experiment), mitochondrial electron transport chain uncoupler CCCP (Sigma, C2759; 20 μM for 2h).

Techniques: Co-Culture Assay, Control, Translocation Assay, Cell Culture, Fluorescence, MANN-WHITNEY