Loss of Function of von Hippel-Lindau Trigger Lipocalin 2-Dependent Inflammatory Responses in Cultured and Primary Renal Tubular Cells

Figure 6

VHL loss of function caused the nuclear translocation of p65 in an LCN2-dependent manner. (a) The HK-2 cells were cultured and treated, as described in Figure 4. Their cell lysates were collected and separated into cytosolic and nuclear fractions. The subcellular localization of NF-κB was detected by Western blot using an antibody against NF-κB’s subunit p65. β-Actin was used as a cytosolic marker and fibrillarin as a nuclear marker. (b) NF-κB’s cytosolic localization was decreased in VHL knockdown (V3) cells; such decrease was attenuated in the VHL (V3) and LCN2 (L1) double knockdown cells. The level of cytosolic p65 (cyto p65) was quantified using ImageJ. (c) NF-κB’s nuclear localization was increased in VHL knockdown (V3) cells; however, such an increase was attenuated in VHL (V3) and LCN2 (L1) double knockdown cells. The expression of nuclear p65 (nuclear p65) was quantified using ImageJ. (d) The HK-2 cells were cultured and treated, as described in Figure 4. The cell lysates were collected and separated into cytosolic and nuclear fractions. The subcellular localization of NF-κB was detected with Western blot using an antibody against the NF-κB subunit p65. β-Actin was used as a cytosolic marker and fibrillarin as a nuclear marker. (e) NF-κB’s cytosolic localization was decreased in VHL knockdown (V1) cells; however, the decrease was attenuated in VHL (V1) and LCN2 (L1) double knockdown cells. The level of cytosolic p65 (cyto p65) was quantified using ImageJ. (f) NF-κB’s nuclear localization was increased in VHL knockdown (V1) cells; the increase was attenuated in VHL (V1) and LCN2 (L1) double knockdown cells. The level of nuclear p65 (nuclear p65) was quantified using ImageJ. All data are presented as the . and .

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