Fig. 3

Immunofluorescence staining for CD68, TNF-α, IL-6, MCP-1, CCR-2 and CCR-5 in the biopsy samples of study subjects. Dual fluorescence staining was done using anti-human CD68 (Alexa 488-Green) and TNF-α, IL-6, MCP-1, CCR-2 and CCR-5 (Alexa 594-red) antibodies to co-localize CD68 with TNF-α, IL-6, MCP-1, CCR-2 and CCR-5 respectively, counterstained with DAPI. Negative controls were run by using isotypes for each fluorochrome. A Immunofluorescence staining in omentum biopsy samples for co-localization of CD68 and TNF-α (Aa, Ab, Ac), co-localization of CD68 and IL-6 (Ad, Ae, Af), co-localization of CD68 and MCP-1 (Ag, Ah, Ai), co-localization of CD68 and CCR-2 (Aj, Ak, Al) and co-localization of CD68 and CCR-5 (Am, An, Ao). B Immunofluorescence staining in subcutaneous biopsy samples for co-localization of CD68 and TNF-α (Ba, Bb, Bc), co-localization of CD68 and IL-6 (Bd, Be, Bf), co-localization of CD68 and MCP-1 (Bg, Bh, Bi), co-localization of CD68 and CCR-2 (Bj, Bk, Bl) and co-localization of CD68 and CCR-5 (Bm, Bn, Bo). C Immunofluorescence staining in liver biopsy samples for co-localization of CD68 and TNF-α (Ca, Cb, Cc), co-localization of CD68 and IL-6 (Cd, Ce, Cf), co-localization of CD68 and MCP-1 (Cg, Ch, Ci), co-localization of CD68 and CCR-2 (Cj, Ck, Cl) and co-localization of CD68 and CCR-5 (Cm, Cn, Co). We found significantly increased immune reactivity of CD68, TNF-α, IL-6, MCP-1, CCR-2 and CCR-5 in omentum, subcutaneous and liver biopsy samples of obese diabetics compared to obese non-diabetics and non-obese subjects (N = 5 non-obese; 16 obese non-diabetics; 17 obese diabetics)