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. 2021 Oct 1;9:727972. doi: 10.3389/fcell.2021.727972

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Copyright © 2021 Shaheen, Akhtar, Umer, Khan, Bakhtiari, Saleem, Faisal and Tariq.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Depletion of chaperonin containing TCP-1 subunit 7 (CCT7) enhances the expression of Polycomb (Pc) group (PcG) target genes. (A–C) Enhanced levels of Abd-B expression observed in CCT7 mutants compared with w¹¹¹⁸. Stage 15 embryos of w¹¹¹⁸ (A) and homozygous mutant embryos for CCT7^KG01477 (B) and CCT7^KG09501 (C) were stained with Abd-B antibody. Homozygous CCT7 embryos (B,C) showed increased Abd-B expression (parasegments marked as PS7-PS14) as compared to w¹¹¹⁸ embryos (A) used as control. All embryos are oriented with their anterior ends upward. (D) As compared to LacZ dsRNA-treated cells, knockdown of CCT7 showed drastic reduction in mRNA levels of CCT7 in D.Mel-2 cells. (E) Significantly increased levels of psq, Antp, Abd-B, Dfd, and Ubx expression determined in CCT7 knockdown cells (D) using real-time PCR compared with LacZ dsRNA-treated cells. Error bars represent SEM for two independent experiments. Statistical significance was calculated using two-way ANOVA (* p ≤ 0.05, **** p ≤ 0.0001). (F–H) Eye-specific knockdown of CCT7 shows a range of homeotic phenotypes, i.e., transformation of eye to duplicated antenna (marked with asterisks, G) and appearance of a leg-like appendage in addition to reduction in eye size (marked with arrow, H). (I) Pie chart representing the frequency of adult phenotypes resulting from eye-specific knockdown of CCT7. A total of 110 adult flies were observed under a stereo microscope (Nikon, C-DSS230) using white light and scored for aberrant eye phenotypes. Phenotypes of adult flies obtained as a result of knockdown of CCT7 were categorized as follows: normal eye (13.64%), reduced eye (70.91%), loss of eye (5.45%), eye to antennal transformation (6.36%), and appearance of rudimentary leg in eye (3.64%). (J) Eye-specific knockdown of CCT7 using a different and stronger RNAi line resulted in 100% lethality of the progeny at the pupal stage (right) as compared to w¹¹¹⁸ control (left). Headless pupal lethal pharate adult observed upon depletion of CCT7 in the eye dissected out from the pupal case (right) as compared to w¹¹¹⁸ control (left).