Fig 1.
CLEC5A knockdown protects HK-2 cells from LPS-induced injury. (A, B) Detection of CLEC5A expression in HK-2 cells by Western blotting. (C) CCK-8 assay for cell viability. (D) Measure LDH production using a commercially available LDH release assay kit. Values are presented as mean ± SD. **p < 0.01, ***p < 0.001. n = 3. CCK-8, cell counting kit-8; CLEC5A, C-type lectin domain family 5 member A; GAPDH, LDH, lactate dehydrogenase; LPS, lipopolysaccharide; SD, standard deviation; shNC, shcontrol.
Fig 2.
CLEC5A knockdown improves HK-2 cells from LPS-induced inflammation. ELISA detects TNF-α (A), IL-6 (B), IL-1β, and (C) levels in HK-2 cells culture fluid. Values are presented as mean ± SD. **p < 0.01, ***p < 0.001. n = 3. CLEC5A, C-type lectin domain family 5 member A; ELISA, enzyme-linked immunosorbent assay; IL, interleukin; LPS, lipopolysaccharide; SD, shNC, TNF-α, tumor necrosis factor-α.
Fig 3.
CLEC5A knockdown improves HK-2 cells from LPS-induced oxidative stress. MDA (A), SOD (B), and GSH-Px (C) levels were measured using commercially available kits in HK-2 cells. Values are presented as mean ± SD. **p < 0.01, ***p < 0.001. n = 3. CLEC5A, C-type lectin domain family 5 member A; GSH-Px, glutathione peroxidase; LPS, lipopolysaccharide; MDA, malondialdehyde; SD, shNC, SOD, superoxide dismutase.
Fig 4.
CLEC5A knockdown can block the activation of NF-κB/NLRP3 signaling pathway. Western blotting to detect p65, p-p65, NLRP3, ASC protein expression. Values are presented as mean ± SD. **p < 0.01, ***p < 0.001. n = 3. ASC, CLEC5A, C-type lectin domain family 5 member A; LPS, lipopolysaccharide; NLRP3, NLR Family, Pyrin Domain Containing Protein 3; SD, shNC.
Fig 5.
Activation of NF-κB reverses the effects of CLEC5A knockdown on LPS-treated HK-2 cells. (A) Western blotting to detect p65, p-p65, NLRP3, and ASC protein expression. (B) CCK-8 assay for cell viability. (C) Measure LDH production using a commercially available LDH release assay kit. (D) ELISA detects IL-1β, IL-6, and TNF-α levels. (E) MDA, SOD, and GSH-Px levels were measured using commercially available kits in HK-2 cells. Values are presented as mean ± SD. **p < 0.01, ***p < 0.001. n = 3. ASC, CCK-8, cell counting kit-8; CLEC5A, C-type lectin domain family 5 member A; ELISA, enzyme-linked immunosorbent assay; GSH-Px, glutathione peroxidase; IL, interleukin; LDH, lactate dehydrogenase; LPS, lipopolysaccharide; MDA, malondialdehyde; NF-κB, NLRP3, NLR Family, Pyrin Domain Containing Protein 3; SD, shNC, SOD, superoxide dismutase; TNF-α, tumor necrosis factor-α.