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. Author manuscript; available in PMC: 2023 Nov 30.
Published in final edited form as: J Mol Biol. 2022 Sep 19;434(22):167832. doi: 10.1016/j.jmb.2022.167832

Figure 4. SRP54 depletion affects expression of membrane proteins with multiple transmembrane domains.

Figure 4.

HeLa Tet-ON cells were transfected with SRP54 specific siRNA (SRP54 KD). Samples for mRNA and protein analysis were prepared as described on Figure 3.

(A) Relative mRNA levels of CHRND and KCNQ1 proteins determined by Deep RNA-seq analysis, shown as a ratio between transcript counts in SRP54 KD and control cells (marked by red dashed line).

(B) mRNA levels of multi-pass membrane proteins were determined by RT-qPCR at 24, 48 and 72 hours after siSRP54 transfection. Data were normalized to actin mRNA and presented relatively to the mRNA levels in control cells (marked by red dashed line). Two-tailed unpaired t test was applied for evaluating statistical significance for 72 hours-time points: ns, not significant; ****, p<0.0001.

(C) Western blot detection of CHRND and KCNQ1 in control (−) and siSRP54 treated (+) cells at different time points after siRNA transfection as indicated. Actin blot was done as a loading control. Numbers represent relative protein expression at 72 h-time point calculated by normalizing the signal in SRP54 depleted cells to the one in the control samples. Quantification of 4–5 biological repeats was done using ImageJ. Standard deviations are shown. Statistical analysis for 72 hours-time point was done using unpaired two-tailed t test. P-values are shown: ns, not significant; **, p < 0.01; ***, p < 0.001.