Fig. 7: Attenuated calcium release responses of Brn-3b KO VSMC is linked to increased proliferative rate.

a (i) qRT-PCR data showing changes in mRNA encoding selected genes (Ryr2, Atp2a1, and Ryr1) in total RNA prepared from primary VSMC cultures that were generated using WT or Brn-3b KO aortas (n = 4–5 aortas per group). (ii) Graph showing quantification of Ca²⁺ transients in primary VSMC cultures from WT or Brn-3b KO aortas, following treatment with 100 µmol ATP. The data shows changes in fluorescent signal intensity over time following treatment with ATP in either WT VSMCs (blue) or Brn-3b KO VSMCs (red). Signals were obtained using confocal microscopy followed by image J analysis with data representing values from six independent WT and Brn-3b KO aortas. (iii) Detailed images showing the significant changes in signal intensity in Brn-3b KO VSMCs, within the first 30 s after ATP treatment, when compared with matched WT controls. To highlight the statistical differences, which was determined using two-way ANOVA. b (i) Graph showing data from BrdU incorporation studies to analyse rate of change in DNA synthesis in primary VSMC cultures from WT or Brn-3b KO aortas. Data represents the mean and standard error of values obtained from seven independent VSMC cultures. (ii) table showing data from independent samples from WT or Brn-3b KO VSMC cultures. (iii) Statistical analysis undertaken using students t test shows significant differences in VSMCs from WT and Brn-3b KO aortas. c Representative fluorescent images of VSMC cultures prepared from either WT or Brn-3b KO aortas and stained with Ki-67 Ab (green) or α-SMA (red). DAPI staining (blue) was used to identify the cell nuclei and merge samples show the overlay of the images as different wavelengths.