Fig. 1: PKP2 in putative fat cells.
From: Impaired Plakophilin-2 in obesity breaks cell cycle dynamics to breed adipocyte senescence
a Pipeline diagram of in vitro cultured human PA growing into differentiated lipid-containing MA. DM-2 (first) and AM-1 (second week) stand for differentiation and adipocyte media, respectively (see in Methods). Expression of PKP2 and adipogenic biomarkers (ADIPOQ, PLIN1, FASN) during differentiation of fat progenitors from (b) SC (n = 4, 3, 3, and 6) and OM (n = 6, 3, 3, and 6 biological replicates for days 0, 2, 7, and 14 of hormonal stimuli, respectively) adipose tissues of the same donor, and from c the SC adipose tissue of one female donor without (BMI < 25 kg/m2) and another one with obesity (BMI >30 kg/m2) (n = 4/timepoint/group). d Impact of macrophage (MM) and macrophage LPS-conditioned media (MCM) on (e) the expression of PKP2 and inflammatory (IL6) and adipogenic (ADIPOQ, FASN) biomarkers in terminally differentiated MA (n = 4/group). f The diagram displays the conditioned differentiation of 3T3-L1 cells into MA, including 4 days under the influence of a differentiation medium (DM), and 4 days of basal preadipocyte culture media (CM) with insulin. Below, the immunofluorescent staining of 3T3-L1 cells exposed to the adipogenic cocktail. Scale bars show sizes of 30 (PA and Day 4) or 20 (MA) µm. g Levels of PKP2 protein in 3T3-L1 cells while developing into MA, as assessed by western blot. The box plot shows the center line at the median, upper and lower lines bound at 75th and 25th percentiles, respectively, and whiskers at minimum and maximum values. h Expression of Pkp2 in differentiating 3T3-L1 cells infected with a retrovirus carrying an empty vector (sh-Scrambled) or sh-Pkp2. i The resulting engineered 3T3-L1 cells in growth conditions showed apparent variations regarding their proliferation rate. j, k Increased amounts of lipids and l, m gene expression patterns were suggestive of enhanced adipogenesis in sh-Pkp2 cells maintained under either j, l non-adipogenic (PA) and k, m differentiating conditions (MA). The scale bar in optical microscope images denotes 100 μm length. Plots for 3T3-L1 cells show data assessed in n = 4 biological replicates/conditions. In all bar plots, results are presented as mean ± SEM. Statistical significance was assessed by two-tailed Fisher’s exact t-test, and by adjusted ANOVA after applying Šidák’s correction to repeated measures (cell growth). r.u. relative units, nd non-detectable, ns non-significant; *p < 0.05; **p < 0.01. Source data are provided as a Source data file.
