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Knock-Down Confirmation of Selected siRNA Targets Using PCR and Western Blot Analysis

This study presents the effective knock-down of specific siRNA targets, including CTSS, CD137L, EFNA3, and others, verified through semi-quantitative PCR and western blot methods. Representative PCR images illustrate the down-regulation of mRNA levels for the indicated targets, with GAPDH serving as an internal control. Additionally, western blot analyses confirm the effectiveness of protein knock-down for targets where antibodies are available, using Tubulin as a control. These results highlight the utility of siRNA in gene silencing applications.

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Knock-Down Confirmation of Selected siRNA Targets Using PCR and Western Blot Analysis

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  1. SelectedsiRNA Control siRNA A B CTSS CTSS SelectedsiRNA Control siRNA 1 0.2 CD137L CD137L 1 0.1 EFNA3 MDK 1 0.2 1 0.2 GDF15 NEO1 1 0.2 1 0.1 GTF2I SERPINI1 1 0.5 1 0.2 MDK PODXL 1 0.1 1 0.3 NEO1 SOSTDC1 1 0.2 1 0.2 SERPINI1 Tubulin 1 0.1 1 0.1 PODXL 1 0.1 1 0.1 SOSTDC1 1 0.1 ZG16B 1 0.1 GAPDH Supplementary Figure S5. Knock-down confirmation by PCR and western blot of the indicated siRNAs targets. A, Representative semi-quantitative PCR images showing the down-regulation at mRNA level of the indicated siRNAs targets. GADPH was used as internal control of the assay. B, Representative western blotting analyses showing the effectiveness of the knock-down at protein level of targets with available antibodies. Tubulin was used as internal control of the assay.

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