Fig. 2.

ORC6 silencing inhibits LPS-induced pro-inflammatory response in PBMCs and BMDMs. Expression levels of both ORC6 and ORC1 in stable peripheral blood mononuclear cells (PBMCs) with the specific ORC6 shRNA (“shORC6-6”) or a scramble control non-sense shRNA (“shC”) were shown (A, B). Cells were exposed to LPS (100 ng/mL) for specified time intervals, mRNA expression and protein secretion (to the medium) of pro-inflammatory cytokines, including IL-1β, TNF-α, and IL-6, were evaluated using qPCR (C-E) and ELISA assays (F-H), respectively. Additionally, assessments of cell viability and death were conducted via CCK-8 assay (I) and medium LDH release (J) assays, respectively. Expression levels of both ORC6 and ORC1 in stable bone marrow-derived macrophages (BMDMs) with the specific murine ORC6 shRNA (“sh-mORC6”) or shC were shown (K and L). The BMDMs were exposed to LPS (100 ng/mL) for specified time intervals, protein secretion (to the medium) of pro-inflammatory cytokines, including IL-1β, TNF-α, and IL-6, were tested (M-O); Cell viability (CCK-8 OD, P) and death (medium LDH release, Q) were tested similarly. The designation “C” denotes vehicle control treatment. Data were presented as mean ± standard deviation (SD, n = 5), with statistical significance indicated by *P < 0.05 compared to “C” and ^#P < 0.05 compared to LPS treatment in “shC” cells. ^#P < 0.05 compared to “shC” (A, B, K and L).Non-statistically significant differences (P > 0.05) were denoted as “N. S.“. These experiments were repeated five times, yielding consistent results across replicates