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(A, B) RAW264.7 cells were infected with ME49 wt or ME49Δ gra3 for 24 h. Cell lysates were analyzed by Western blot for total <t>STAT6</t> and phospho‑STAT6 (p‑STAT6). (C, D) Cells were pretreated with the STING inhibitor H‑151 (1 μM, MCE) for 1 h before infection with the indicated strains for 24 h, followed by Western blot analysis of STAT6, p‑STAT6 and profilin. (E, F) Cells were pretreated with the <t>AS1517499</t> (100 nM, MCE) for 1 h before infection with the indicated strains for 24 h, followed by Western blot analysis of STAT6, p‑STAT6 and profilin. (G) Following the same pretreatment and infection protocol, intracellular parasite replication was assessed by examining at least 200 vacuoles and categorizing them based on the number of parasites per vacuole (2, 4, 8, or >8). Data were presented as the mean ± SD from three independent experiments. Statistical significance is indicated by * p < 0.05, ** p < 0.01 and *** p < 0.001. Data were compared using one-way analysis of variance.
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(A, B) RAW264.7 cells were infected with ME49 wt or ME49Δ gra3 for 24 h. Cell lysates were analyzed by Western blot for total <t>STAT6</t> and phospho‑STAT6 (p‑STAT6). (C, D) Cells were pretreated with the STING inhibitor H‑151 (1 μM, MCE) for 1 h before infection with the indicated strains for 24 h, followed by Western blot analysis of STAT6, p‑STAT6 and profilin. (E, F) Cells were pretreated with the <t>AS1517499</t> (100 nM, MCE) for 1 h before infection with the indicated strains for 24 h, followed by Western blot analysis of STAT6, p‑STAT6 and profilin. (G) Following the same pretreatment and infection protocol, intracellular parasite replication was assessed by examining at least 200 vacuoles and categorizing them based on the number of parasites per vacuole (2, 4, 8, or >8). Data were presented as the mean ± SD from three independent experiments. Statistical significance is indicated by * p < 0.05, ** p < 0.01 and *** p < 0.001. Data were compared using one-way analysis of variance.
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(A, B) RAW264.7 cells were infected with ME49 wt or ME49Δ gra3 for 24 h. Cell lysates were analyzed by Western blot for total <t>STAT6</t> and phospho‑STAT6 (p‑STAT6). (C, D) Cells were pretreated with the STING inhibitor H‑151 (1 μM, MCE) for 1 h before infection with the indicated strains for 24 h, followed by Western blot analysis of STAT6, p‑STAT6 and profilin. (E, F) Cells were pretreated with the <t>AS1517499</t> (100 nM, MCE) for 1 h before infection with the indicated strains for 24 h, followed by Western blot analysis of STAT6, p‑STAT6 and profilin. (G) Following the same pretreatment and infection protocol, intracellular parasite replication was assessed by examining at least 200 vacuoles and categorizing them based on the number of parasites per vacuole (2, 4, 8, or >8). Data were presented as the mean ± SD from three independent experiments. Statistical significance is indicated by * p < 0.05, ** p < 0.01 and *** p < 0.001. Data were compared using one-way analysis of variance.
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https://www.bioz.com/result/stat6 in 1/product/MedChemExpress
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stat6 in 3  (MedChemExpress)
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(A, B) RAW264.7 cells were infected with ME49 wt or ME49Δ gra3 for 24 h. Cell lysates were analyzed by Western blot for total <t>STAT6</t> and phospho‑STAT6 (p‑STAT6). (C, D) Cells were pretreated with the STING inhibitor H‑151 (1 μM, MCE) for 1 h before infection with the indicated strains for 24 h, followed by Western blot analysis of STAT6, p‑STAT6 and profilin. (E, F) Cells were pretreated with the <t>AS1517499</t> (100 nM, MCE) for 1 h before infection with the indicated strains for 24 h, followed by Western blot analysis of STAT6, p‑STAT6 and profilin. (G) Following the same pretreatment and infection protocol, intracellular parasite replication was assessed by examining at least 200 vacuoles and categorizing them based on the number of parasites per vacuole (2, 4, 8, or >8). Data were presented as the mean ± SD from three independent experiments. Statistical significance is indicated by * p < 0.05, ** p < 0.01 and *** p < 0.001. Data were compared using one-way analysis of variance.
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stat6in 3  (MedChemExpress)
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(A, B) RAW264.7 cells were infected with ME49 wt or ME49Δ gra3 for 24 h. Cell lysates were analyzed by Western blot for total <t>STAT6</t> and phospho‑STAT6 (p‑STAT6). (C, D) Cells were pretreated with the STING inhibitor H‑151 (1 μM, MCE) for 1 h before infection with the indicated strains for 24 h, followed by Western blot analysis of STAT6, p‑STAT6 and profilin. (E, F) Cells were pretreated with the <t>AS1517499</t> (100 nM, MCE) for 1 h before infection with the indicated strains for 24 h, followed by Western blot analysis of STAT6, p‑STAT6 and profilin. (G) Following the same pretreatment and infection protocol, intracellular parasite replication was assessed by examining at least 200 vacuoles and categorizing them based on the number of parasites per vacuole (2, 4, 8, or >8). Data were presented as the mean ± SD from three independent experiments. Statistical significance is indicated by * p < 0.05, ** p < 0.01 and *** p < 0.001. Data were compared using one-way analysis of variance.
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(A, B) RAW264.7 cells were infected with ME49 wt or ME49Δ gra3 for 24 h. Cell lysates were analyzed by Western blot for total <t>STAT6</t> and phospho‑STAT6 (p‑STAT6). (C, D) Cells were pretreated with the STING inhibitor H‑151 (1 μM, MCE) for 1 h before infection with the indicated strains for 24 h, followed by Western blot analysis of STAT6, p‑STAT6 and profilin. (E, F) Cells were pretreated with the <t>AS1517499</t> (100 nM, MCE) for 1 h before infection with the indicated strains for 24 h, followed by Western blot analysis of STAT6, p‑STAT6 and profilin. (G) Following the same pretreatment and infection protocol, intracellular parasite replication was assessed by examining at least 200 vacuoles and categorizing them based on the number of parasites per vacuole (2, 4, 8, or >8). Data were presented as the mean ± SD from three independent experiments. Statistical significance is indicated by * p < 0.05, ** p < 0.01 and *** p < 0.001. Data were compared using one-way analysis of variance.
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(A, B) RAW264.7 cells were infected with ME49 wt or ME49Δ gra3 for 24 h. Cell lysates were analyzed by Western blot for total STAT6 and phospho‑STAT6 (p‑STAT6). (C, D) Cells were pretreated with the STING inhibitor H‑151 (1 μM, MCE) for 1 h before infection with the indicated strains for 24 h, followed by Western blot analysis of STAT6, p‑STAT6 and profilin. (E, F) Cells were pretreated with the AS1517499 (100 nM, MCE) for 1 h before infection with the indicated strains for 24 h, followed by Western blot analysis of STAT6, p‑STAT6 and profilin. (G) Following the same pretreatment and infection protocol, intracellular parasite replication was assessed by examining at least 200 vacuoles and categorizing them based on the number of parasites per vacuole (2, 4, 8, or >8). Data were presented as the mean ± SD from three independent experiments. Statistical significance is indicated by * p < 0.05, ** p < 0.01 and *** p < 0.001. Data were compared using one-way analysis of variance.

Journal: PLOS Neglected Tropical Diseases

Article Title: Toxoplasma gondii GRA3 activates interferon-stimulated genes and STAT6 by the cGAS-STING pathway to promote parasite proliferation

doi: 10.1371/journal.pntd.0014035

Figure Lengend Snippet: (A, B) RAW264.7 cells were infected with ME49 wt or ME49Δ gra3 for 24 h. Cell lysates were analyzed by Western blot for total STAT6 and phospho‑STAT6 (p‑STAT6). (C, D) Cells were pretreated with the STING inhibitor H‑151 (1 μM, MCE) for 1 h before infection with the indicated strains for 24 h, followed by Western blot analysis of STAT6, p‑STAT6 and profilin. (E, F) Cells were pretreated with the AS1517499 (100 nM, MCE) for 1 h before infection with the indicated strains for 24 h, followed by Western blot analysis of STAT6, p‑STAT6 and profilin. (G) Following the same pretreatment and infection protocol, intracellular parasite replication was assessed by examining at least 200 vacuoles and categorizing them based on the number of parasites per vacuole (2, 4, 8, or >8). Data were presented as the mean ± SD from three independent experiments. Statistical significance is indicated by * p < 0.05, ** p < 0.01 and *** p < 0.001. Data were compared using one-way analysis of variance.

Article Snippet: Treatment with the STAT6 phosphorylation inhibitor AS1517499 (100 nM, MCE) suppressed GRA3-induced STAT6 activation and T. gondii profilin expression ( and ).

Techniques: Infection, Western Blot

By engaging host STING, the Toxoplasma gondii protein GRA3 activates TBK1 and IRF3, thereby inducing STAT6 phosphorylation and IRF3-mediated expression of IFN-β and ISG56. This regulated immune activation prevents excessive parasite proliferation, thus facilitating the establishment of a stable chronic infection within the host.

Journal: PLOS Neglected Tropical Diseases

Article Title: Toxoplasma gondii GRA3 activates interferon-stimulated genes and STAT6 by the cGAS-STING pathway to promote parasite proliferation

doi: 10.1371/journal.pntd.0014035

Figure Lengend Snippet: By engaging host STING, the Toxoplasma gondii protein GRA3 activates TBK1 and IRF3, thereby inducing STAT6 phosphorylation and IRF3-mediated expression of IFN-β and ISG56. This regulated immune activation prevents excessive parasite proliferation, thus facilitating the establishment of a stable chronic infection within the host.

Article Snippet: Treatment with the STAT6 phosphorylation inhibitor AS1517499 (100 nM, MCE) suppressed GRA3-induced STAT6 activation and T. gondii profilin expression ( and ).

Techniques: Phospho-proteomics, Expressing, Activation Assay, Infection