Figure 4
Differential enrichment of Mup transcripts in the SC WAT depot. A. qPCR assessment of transcript levels in SVF and AD fractions of SC and EP WAT using the Mup1 primer set. SVF, stromal vascular fraction; AD, adipocyte fraction. B. qPCR assessment of transcript levels in SVF and AD of SC and EP WAT using the Mup1/2 primer set. C. qPCR assessment of whole SC, EP, RP or BAT adipose tissue using the Mup1 primer set. D. qPCR assessment of whole SC, EP, RP or BAT adipose tissue using the Mup1/2 primer set. For A the EP SVF signal level was set to a value of 1 and for B the EP AF signal was set to 1. For A, * indicates p < 0.001 for SC SVF vs. EP SVF and for SC AD vs. EP AD, and # indicates p < 0.01 for SC SVF vs. all others. For B, * indicates p < 0.001 for SC SVF vs. EP SVF and for SC AD vs. EP AD, and # indicates p < 0.001 for SC AD vs. all others. For C and D the signal level in BAT was set to 1 and * indicates p < 0.001 for SC or RP compared with EP and with BAT. E. Upper panel shows Northern blot analysis of depot-dependent gene expression in either SC WAT or EP WAT depots of four individual male C57Bl/6J mice using 32P dATP-labeled Mup1, aFABP or SCD1 hybridization probes. Lanes 1–4 and lanes 5–8 represent SC and EP WAT tissue from mouse #1, #2, #3 and #4, respectively. Lower panel shows Northern blot analysis of fractionated SC and EP WAT. SV, stromal vascular fraction; AF, adipocyte fraction. Ethidium bromide staining of rRNA is shown as gel loading control. F. and G. qPCR assessment for 3T3-L1 or primary cultures of preadipocytes (Pre) and adipocytes (Adi) using the Mup1 (F) or the Mup1/2 (G) primer sets. The level of transcript expression in primary preadipocytes was set to a value of 1. For F and G, * indicates p < 0.001 for primary adipocytes vs. all others. For A-D, F and G, the left panels show data corrected against Gapdh and the right panels show data corrected against 36B4 as internal control for qPCR analysis; values stated in the text are the average of the Gapdh and 36B4-corrected data for each comparison.