Fig. 1

Analysis of hnRNPD expression in oral tissues. a Quantitative real time PCR (qPCR) analysis of hnRNPD transcripts in oral cancer cells, OSCCs and normal tissues. i Analysis of hnRNPD mRNA levels in oral cancer cell line. Total cellular RNA from various oral cancer cell lines (SCC-4, HSC-2, Tu-167, MDA-1986) were isolated, reverse transcribed and subjected to real time PCR analysis using specific hnRNPD primers. Simultaneously real time PCR for 18S ribosomal RNA was performed and served as an internal control for normalization. After real time PCR the 2^−Δct value of hnRNPD mRNA obtained in SCC-4 cells was assigned a value of 1 and the fold increase in other cell lines was calculated relative to this. Histogram shows the relative abundance of hnRNPD transcripts in SCC4, HSC2, Tu167 and MDA1986. ii Expression of hnRNPD transcripts in 12 random OSCCs samples and their paired normal tissues. Total cellular RNA was isolated from OSCCs and their paired normal tissues, reverse transcribed, and subjected to real time PCR analysis using specific hnRNPD primers. Parallel real time PCR for 18S ribosomal RNA was performed which served as an internal control for normalization of hnRNPD mRNA levels in OSCC and paired normal tissues. After normalization the fold increase in the levels of hnRNPD transcripts in each OSCC sample over their respective paired normal tissue was calculated and plotted individually. hnRNPD mRNA levels were significantly higher in oral OSCCs patients as compared to their paired normal oral tissue samples (p = 0.007, Mann–Whitney U test). Bar graph data is represented by fold changes of OSCCs samples after normalization with paired normal tissue samples. iii A representation of scatter plot after normalized 2^−Δct values of each OSCCs and paired normal tissue samples. b Western blot analysis. Photomicrograph showing protein expression of hnRNPD in (i) oral cancer cell lines (SCC4, HSC2, Tu167 and MDA1986); ii tissue lysates obtained from normal oral mucosa (n1–n4), oral dysplasia (d1, d2) and OSCCs (t1–t8). Panel shows increased expression of hnRNPD in dysplasia (d1, d2) and OSCCs (t1–t8) as compared to normal mucosa (n1–n4). Whole cell lysates prepared from SCC4 cells was used as a positive control. β-actin was used as loading control (lower panel)