Dear Sofia, please excuse the misunderstandings caused by my terminology. You can rename all lowercase "fc" below to normalized_signal or log transformed ratio of signal to input. Only the 2 new "FC" are log2foldchanges. At http://genomics-lab.fleming.gr/fleming/DKlab/run392/run402/exonGencode.groups.rpkm-v2.csv you can load the RPKM analysis per exon for the liver samples with corrected q-values (FDR). With a FDR <= 0.1 (which is common practice), there are 804 exons for M2RIP_WTvsInputWT_vs_M2RIP_Tg6105vsInputTg6105 and 73 exons for HuR_RIPC57r_vs_IgG_RIPC57 with a significant signal. I've also added the foldchanges in the last 2 columns. The definition of the exon in col. 1 to 6: [1] "Geneid" [2] "Chr" [3] "Start" [4] "End" [5] "Strand" [6] "Length" The non-input RPKM values in col 7 to 18: [7] "DKR13r_M2RiPWt_1M" [8] "DKR14r_M2RiPWt_2M" [9] "DKR15r_M2RiPWt_3F" [10] "DKR16_M2RiPTg6105_1M" [11] "DKR17r_M2RiPTg6105_2M" [12] "DKR18_M2RiPTg6105_3F" [13] "DKR19_HuRRiPC57_1M" [14] "DKR21_HuRRiPC57_3F" [15] "DKR20_HURRIPC57_2M" [16] "DKR22_IgGRiPC57_1M" [17] "DKR23r_IgGRiPC57_2M" [18] "DKR24r_IgGRiPC57_3F" The input RPKM values (averaged over the 3 replicates) in col 19 to 21 [19] "iWT" [20] "iTg6105" [21] "iC57" The log2 foldchanges as defined below in (a) in col 22 to 30 [22] "M2RIP_WT1_fc" [23] "M2RIP_WT2_fc" [24] "M2RIP_WT3_fc" [25] "M2RIP_Tg6105_1_fc" [26] "M2RIP_Tg6105_2_fc" [27] "M2RIP_Tg6105_3_fc" [28] "HuR_RIPC57_1_fc" [29] "HuR_RIPC57_2_fc" [30] "HuR_RIPC57_3_fc" p-values and correced p-vaules (FDR or q-values) of a t.test for M2RIP_WTvsInputWT vs M2RIP_Tg6105vsInputTg610 [31] "M2RIP_WTvsInputWT_vs_M2RIP_Tg6105vsInputTg6105_pval" [32] "M2RIP_WTvsInputWT_vs_M2RIP_Tg6105vsInputTg6105_qval" The log2 foldchanges as defined below in (b) [33] "IgG_RIPC57_1_fc" [34] "IgG_RIPC57_2_fc" [35] "IgG_RIPC57_3_fc" p-values and correced p-vaules (FDR or q-values) of a t.test for HuR_RIPC57rVSinput_vs_IgG_RIPC57VSinput [36] "HuR_RIPC57r_vs_IgG_RIPC57_pval" [37] "HuR_RIPC57r_vs_IgG_RIPC57_qval" log2 fold changes of normalized signals, calculated using (c): [38] "M2RIP_WTvsInputWT_vs_M2RIP_Tg6105vsInputTg6105_FC" [39] "HuR_RIPC57r_vs_IgG_RIPC57_FC" (a) # M2RIP_WT vs InputWT r$M2RIP_WT1_fc=log2((1+r$DKR13r_M2RiPWt_1M)/(1+r$iWT)); r$M2RIP_WT2_fc=log2((1+r$DKR14r_M2RiPWt_2M)/(1+r$iWT)); r$M2RIP_WT3_fc=log2((1+r$DKR15r_M2RiPWt_3F)/(1+r$iWT)); # M2RIP_Tg6105 vs InputTg6105 r$M2RIP_Tg6105_1_fc=log2((1+r$DKR16_M2RiPTg6105_1M )/(1+r$iTg6105)); r$M2RIP_Tg6105_2_fc=log2((1+r$DKR17r_M2RiPTg6105_2M)/(1+r$iTg6105)); r$M2RIP_Tg6105_3_fc=log2((1+r$DKR18_M2RiPTg6105_3F )/(1+r$iTg6105)); # HuR_RIPC57 vs InputC57 r$HuR_RIPC57_1_fc=log2((1+r$DKR19_HuRRiPC57_1M)/(1+r$iC57)); r$HuR_RIPC57_2_fc=log2((1+r$DKR21_HuRRiPC57_3F)/(1+r$iC57)); r$HuR_RIPC57_3_fc=log2((1+r$DKR20_HURRIPC57_2M)/(1+r$iC57)); (b) r$IgG_RIPC57_1_fc=log2((1+r$DKR22_IgGRiPC57_1M)/(1+r$iC57)); r$IgG_RIPC57_2_fc=log2((1+r$DKR23r_IgGRiPC57_2M)/(1+r$iC57)); r$IgG_RIPC57_3_fc=log2((1+r$DKR24r_IgGRiPC57_3F)/(1+r$iC57)); (c) a=apply(cbind(r$M2RIP_WT1_fc,r$M2RIP_WT2_fc,r$M2RIP_WT3_fc),1,mean) b=apply(cbind(r$M2RIP_Tg6105_1_fc,r$M2RIP_Tg6105_2_fc,r$M2RIP_Tg6105_3_fc),1,mean) r$M2RIP_WTvsInputWT_vs_M2RIP_Tg6105vsInputTg6105_FC=a-b a=apply(cbind(r$HuR_RIPC57_1_fc,r$HuR_RIPC57_2_fc,r$HuR_RIPC57_3_fc),1,mean) b=apply(cbind(r$IgG_RIPC57_1_fc,r$IgG_RIPC57_2_fc,r$IgG_RIPC57_3_fc),1,mean) r$HuR_RIPC57r_vs_IgG_RIPC57_FC=a-b BW, Martin