Fig. 4

In vitro effects of Pioglitazone treatment in non-diabetic splenocytes. a Left, Similar levels of PPARγ in splenocytes with and without Pioglitazone treatment. Right, Increase in PPARγ levels in Pio-treated hepatocytes compared to untreated. Splenocytes or hepatocytes were cultured in the presence or absence of Pioglitazone 25 μM for 24 h followed by cell lysis treatment for Western blot analysis. Results are shown as bar graphs, dark gray represent untreated, white with black stripes represent Pio-treated. Graphs show the ratio of PPARγ to β-actin as arbitrary units (A.U.); n = 3. Upper part shows blot results in presence (+Pio) or absence (untreated) of treatment. b Top panels show the percentage of CD3⁺ and CD3⁺CD4⁺ T lymphocytes in untreated and Pio-treated splenocytes; n = 3. Cultured splenocytes were harvested and surface labeled with anti-CD3 Alexa Fluor 647, -CD4 PE-Cy7 and -CD25 FITC antibodies prior fixation and permeabilization for intracellular FoxP3-PE labeling. Mid panel shows percentage of CD3⁺CD4⁺CD25⁺ T cells followed by FoxP3 analysis of the gated population. Histogram shows percentage of FoxP3⁺ cells from the CD3⁺CD4⁺CD25⁺ gate; n = 3. Gray histogram depicts isotype control; black line depicts anti-rat FoxP3 labeling. Figures are representation of n = 3. At the right there is quantitation of each panel in bar graph: *p = 0.0006, ^# p = 0.002, ^¶ p = 0.0237