Figure 2: The tetracycline COL-3 reduces the
inflammatory response of microglial cells challenged with either LPS or
αSa (A) (b) TEM images showing the presence of αS amyloid fibrils under
shaken conditions, only (lower image). Scale bar: 1 μm. (B)Quantification of TNF-α levels in microglial cells exposed or not to LPS
(10 ng/ml, 24-h) in the presence or not of COL-3 (1, 10 or 20 μM), DOX
(50µM) or the reference antiinflammatory drug DEX (2.5 μM). Data are
means ± SEM (n = 4), *p < 0.05 vs control.#p < 0.05 vs LPS. One-way ANOVA
followed by Bonferroni post-hoc test. (C) Quantification of
Iba-1 immunofluorescence density in microglial cultures exposed or not
to to LPS (10 ng/ml, 24-h) in the presence or not of COL-3 (10 or 20
μM), DOX (50µM) or the reference glucocorticoid DEX (2.5 μM). Data are
means ± SEM (n = 4), *p < 0.05 vs control.#p < 0.05 vs LPS. One-way ANOVA
followed by Bonferroni post-hoc test. (D) Visualization of
CD11b (green) and Iba-1 (red) immunofluorescence signals of LPS (10
ng/ml)-treated microglial cell cultures treated or not with COL-3 (20µM)
or DOX (50 µM). Cell nuclei were counterstained with Hoechst 33342
(blue). Scale bar = 120 µm. (E) Quantification of TNF-α levels
in microglial cells exposed or not to αSa
(70 µg/ml, 24-h) in the presence or
not of COL-3 (10 and 20 μM), DOX (50µM) or DEX (2.5 μM). Some cultures
were also exposed αSm (70 µg/ml). Data are means ± SEM (n = 4), *p
< 0.05 vs control. #p <
0.05 vs αSa. One-way ANOVA followed by Bonferroni post-hoc test.(F) Quantification of Iba-1 immunofluorescence signals in
microglial cultures exposed or not to to α Sa (70 µg/ml, 24-h) in the
presence or not of COL-3 (10 or 20 μM), DOX (50µM) or DEX (2.5 μM). Data
are means ± SEM (n = 4), *p < 0.05 vs control.#p < 0.05 vs αSa. One-way ANOVA
followed by Bonferroni post-hoc test. (G) Visualization of
CD11b (green) and Iba-1 (red) immunofluorescent signals in αSa (70
µg/ml)-treated microglial cell cultures exposed or not to COL-3 (20µM)
or DOX (50 µM). Cell nuclei were counterstained with Hoechst 33342
(blue). Scale bar = 120 µm.