Fig. 3

IL-1β induces the expression of HSP60 in vitro, in vivo, and in primary microglial culture. a, b Left panel depicts the representative western blot image showing increase in the level of HSP60 at different time points after IL-1β treatment in N9 cells (in vitro) (a) and in BALB/c mice brains (in vivo) (b), respectively. Right panel represents the bar diagrams which depict mean fold change in the levels of HSP60 with respect to control treated group. Thirty microgram protein was loaded for western blot, and β-actin served as a loading control. c, d Quantitative real-time PCR analysis of the transcript level of HSP60 after treatment with IL-1β at different time points in N9 murine microglial cells (c) and in BALB/c mice brain (d). GAPDH was used for the normalization. e, f Immunostaining of HSP60 in primary microglia (e) and N9 murine microglial cells (f) using specific antibodies as described in methods. Nuclei were counterstained with the DNA-binding dye DAPI. Images were captured using Zeiss apotome fluorescence microscope (Scale bar—20 μm; magnification—×40). Representative of three independent experiments is shown here (a, b, e, f) (n = 3). *p < 0.05, **p < 0.01 in comparison to control values. Data represented are mean ± SD of three independent experiments