Dysregulated Alternative Splicing Pattern of PKC𝜹 during Differentiation of Human Preadipocytes Represents Distinct Differences between Lean and Obese Adipocytes

Figure 3

(a) Western blot analysis of adipose tissue lysates obtained from obese depots from (1) omental; (2) subcutaneous. The blots are representative of experiments repeated thrice from the lysates. Graph represents percent PKCδ exon inclusion calculated as PKC𝛿VIII/(PKC𝛿VIII + PKC𝛿I) × 100 and is representative of three experiments. (b) Human lean and obese preadipocytes were differentiated in vitro. Cells were serum deprived for 48 h on day 0 (preadipocytes) and on day 10 (mature adipocytes). Annexin V/PI staining was analyzed by flow cytometry. The first plot demonstrates the gating of the adipocyte population. The remaining plots shown are lean control, lean-serum deprived, obese control, and obese serum-deprived and are divided into four quadrants based on Annexin V (AV) and propidium iodide staining (PI) staining. Q1-LL represents viable (unstained) cells. Q1-LR represents AV staining only (early apoptosis). Q1-UR represents cells stained with both AV and PI (later apoptosis). Q1-UL represents cells stained with PI only (end-stage apoptosis). Data shown are representative of five different experiments. (c) Graph represents AV/PI and represents 5 experiments. Statistical analysis performed by two-way ANOVA; 𝑃>0.75 ns, not significant within group; ***𝑃<0.0001 highly significant between control and serum-deprived; ***𝑃<0.0001 highly significant between lean and obese; ***𝑃<0.0001 highly significant between days 0 and 10 serum-deprived.