Fig 1: Host SUMOylation promotes the growth of Leishmania donovani by suppressing pro-inflammatory cytokines. THP-1 macrophages were transfected with specific siRNAs followed by the infection of L. donovani along with or without the stimulation of LPS (100 ng/ml) for 24 h MOCK here represents the transfection with a control siRNA. Culture supernatants were collected to measure different cytokines, while cells were used to check the expression of inflammatory markers at the transcript level. (A) Graph represents cytokine levels measured by sandwich ELISA. Statistical significance was quantified using the unpaired t-test with Welch’s correction, *p = 0.012 for MOCK vs. AOS-1 and ***p = 0.0003 for MOCK vs. SENP1 for IL-10; **p = 0.0029, 0.0032, 0.0028, and 0.0074 for MOCK vs. SUMO-1, AOS-1, UBA2, and SENP1, respectively, and ***p = 0.0002 for MOCK vs. SUMO-2/3 for IL-12p40; *p = 0.025 for MOCK vs. SUMO-2/3, **p = 0.002 for MOCK vs. SENP1, and ***p = 0.0002 and 0.0008 for MOCK vs. UBA2 and UBC9, respectively, for IFN-?; *p = 0.049 for MOCK vs. UBC9, **p = 0.0012 for MOCK vs. AOS-1, ****p < 0.0001 for SUMO-2/3 and UBA2 for TNF-a in LPS-stimulated and Ld-infected macrophages. (B) RNA was isolated for gene expression analysis of inflammatory cytokines by quantitative real-time PCR (qRT-PCR). *p = 0.04 for MOCK vs. SUMO-2/3 or UBC9 for IL-10; *p = 0.04 and 0.05 for MOCK vs. UBA2 and SENP1, respectively; **p = 0.008 for MOCK vs. UBC9 for IL-12; *p = 0.02 for MOCK vs. SUMO-2/3 and 0.04 for MOCK vs. SUMO-1 or UBA2 or UBC9 for IL-32?; *p = 0.05 and 0.02 for MOCK vs. UBA2 and SENP1, respectively; **p = 0.01 and 0.008 for MOCK vs. SUMO-1 and SUMO-2/3 for TNF-a in LPS-stimulated and Ld-infected macrophages. p-Value was calculated based on Student’s unpaired 2-tailed t-test. LPS, lipopolysaccharide.
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