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. 2023 Jun 15;37(7):e23008. doi: 10.1096/fj.202300100R

FIGURE 3.

FIGURE 3

siRNA‐based cystin depletion leads to FPC reduction. (A) siRNA depletion of Cys1 mRNA. The efficiency of siRNA oligos (60 nM) to reduce Cys1 mRNA levels was measured relative to Gapdh1 mRNA in wt, wt + siRNA scrambled, and wt + siRNA Cys1 samples. Significant (**p < .0001, unpaired t‐test) reduction in Cys1 mRNA levels confirmed the specificity of the selected oligos. (B) siRNA depletion of Cys1 mRNA leads to reduction in FPC and cystin protein levels. Representative WB of FPC (top) and cystin (bottom) following cystin siRNA treatment for 72 h (siRNA concentrations indicated above image). GAPDH levels (middle) were used to normalize cystin and FPC expression. FPC expression was tested using anti‐FPC C‐terminal, rat monoclonal antibody, cystin expression was measured using anti‐cystin rabbit polyclonal antibody (bottom). (C) Densitometric analysis of cystin protein depletion (left) and corresponding FPC levels in the same samples (right). Values are plotted relative to GAPDH protein from N = 1–3 separate experiments, with n = 3–6 under each siRNA concentration (****p < .0001, unpaired t‐test, significance was determined between scrambled and cystin‐specific siRNA (80 nM)).