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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 3. SRP54 depletion affects secreted proteins expression.

Figure 3.

HeLa Tet-ON cells were transfected with SRP54 specific siRNA (siSRP54). Samples for mRNA and protein analysis were collected at 24, 48 and 72 hours after transfection.

(A) Relative mRNA levels determined by Deep RNA-seq analysis, shown as a ratio between transcript counts in SRP54 KD and control cells.

(B) mRNA levels of secreted proteins, SRP54 and the control cytosolic protein HPRT in SRP54 KD cells determined by RT-qPCR in independent from the Deep RNA-seq set of experiments, 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 lines). Standard errors were calculated based on the values from three independent experiments. 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 CD248, PDIA3, SRP54 and actin in control (−) and siSRP54 treated (+) cells at different time points after siRNA transfection as indicated. Actin blot was done as a loading control.

(D) Immunostaining of control and SRP knockdown (SRP54 KD) cells with CD248 (top), PDIA3 (middle) and SRP54 (bottom) specific antibodies. Cells were stained at 48 hours after siSRP54 transfection.