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. 2018 Nov 19;14(11):e1007437. doi: 10.1371/journal.ppat.1007437

Fig 3. Effect of SIRT2 inhibition is nitric oxide mediated.

Fig 3

A. Nitric oxide levels in conditioned media in response to SIRT2 inhibition at indicated time post infection. (UI- uninfected DCs, UI AK7- Uninfected DCs treated with 10 μM AK7, STM- Salmonella Typhimurium infected DCs, STM AK7- - Salmonella Typhimurium infected DCs treated with 10 μM AK7) (Data are presented as mean ± SEM of 5 independent experiments). B. qPCR analysis of NOS2 expression in DCs at indicated time post infection in response to SIRT2 inhibition. GAPDH was used as an internal control. (UI- uninfected DCs, UI AK7- Uninfected DCs treated with 10 μM AK7, STM- Salmonella Typhimurium infected DCs, STM AK7- Salmonella Typhimurium infected DCs treated with 10 μM AK7, LPS- 100 ng/ml LPS treated). (Data are presented as mean ± SEM of 3 independent experiments). C. Representative immunoblot of NOS2 in the presence and absence of SIRT2 inhibition at indicated time. (UI- uninfected DCs, UI AK7- Uninfected DCs treated with 10 μM AK7, STM- Salmonella Typhimurium infected DCs, STM AK7- Salmonella Typhimurium infected DCs treated with 10 μM AK7) D. Densitometry analysis of NOS2 level in the presence and absence of SIRT2 inhibition at indicated time. (UI- uninfected DCs, UI AK7- Uninfected DCs treated with 10 μM AK7, STM- Salmonella Typhimurium infected DCs, STM AK7- Salmonella Typhimurium infected DCs treated with 10 μM AK7) (Data are presented as mean ± SEM of 3 independent experiments). E. Percentage survival of intracellular Salmonella in DCs in gentamicin protection assay in the presence and absence of SIRT2 inhibitor in wild type and NOS2-/- DCs. (Mock- DMSO treated, AK7- 10 μM AK7 treated). (Data are presented as mean ± SEM of 4 independent experiments) F. 3[H] Thymidine incorporation assay to assess CD8+ T cells proliferation during Salmonella infection in wild type and NOS2-/- DCs in the presence and absence of SIRT2 inhibition. (Ova- Ovalbumin, DC- Dendritic cells, T cell- mixed lymphocyte, STM- Salmonella Typhimurium, ConA- Concanavalin A) (Data are presented as mean ± SEM of 3 independent experiments) G. IL-2 levels during CD8+T cells proliferation assay in response to Salmonella infection in wild type and NOS2-/- DCs. (Ova- Ovalbumin, DC- Dendritic cells, T cell- mixed lymphocyte, STM- Salmonella Typhimurium) (Data are presented as mean ± SEM of 2 independent experiments) (unpaired two tailed Student’s t- test, p- value, *** < 0.0001, **<0.001, *<0.01).